Specification

The variety of life, both past and present, is extensive, but the biochemical basis of life is similar for
all living things.

Monomers are the smaller units from which larger molecules are made.

Polymers are molecules made from a large number of monomers joined together.

Monosaccharides, amino acids and nucleotides are examples of monomers.

A condensation reaction joins two molecules together with the formation of a chemical bond and
involves the elimination of a molecule of water.

A hydrolysis reaction breaks a chemical bond between two molecules and involves the use of a
water molecule.

Previous knowledge

How to test for starch?

How to test for protein?
How to test for glucose?
What is starch made up from?
What are proteins made up from?
What are fats made up from?
What is a polymer?

Biofact sheets: 39, 42, 74, 80, 152, 159, 173

Monomers are the smaller units from which larger molecules are made. Key monomers to learn Isomers: molecules with the same molecular formula but
different molecular arrnagement
Nucleic acids, amino acids, & glucose, fructose, fatty acids and glycerol

DNA nucleotide glucose Fructose

glucose
Amino Acid

RNA nucleotide

Glycerol

Fatty Acid
Polymers: long chains of repeating monomers joined by condensation reactions. Water is Hydrolysis: reaction that breaks chemical bonds and involves the
removed in a condensation reaction and these polymers form by condensation polymerisation use of water

Proteins are joined by peptide bonds

Carbohydrates are joined by glycosidic bonds

This particular reaction is producing a disaccharide. There are three common

The molecular formula of disaccharides produced
fructose is C6H12O6. What is
the molecular formula of Maltose: glucose + glucose
sucrose?
Sucrose: glucose + fructose
C12;
H22O11; Lactose: glucose + galactose
Triglyceride formation:

Triglycerides are not polymers as they are not made up of long chains of repeating subunits. But they are macromolecules
Condensation polymerisation in DNA formation occurs between the
phosphate of one nucleotide and the pentose sugar of the other. A
phosphodiester bond is produced.
A closer look at carbohydrates
Specification
Monosaccharides are the monomers from which larger carbohydrates are made. Glucose, galactose
and fructose are common monosaccharides.

A condensation reaction between two monosaccharides forms a glycosidic bond.

Disaccharides are formed by the condensation of two monosaccharides:

Maltose is a disaccharide formed by condensation of two glucose molecules
Sucrose is a disaccharide formed by condensation of a glucose molecule and a fructose molecule
Lactose is a disaccharide formed by condensation of a glucose molecule and a galactose molecule.

Glucose has two isomers, -glucose and -glucose, with structures

Polysaccharides are formed by the condensation of many glucose units.

Glycogen and starch are formed by the condensation of -glucose.
Cellulose is formed by the condensation of -glucose.

The basic structure and functions of glycogen, starch and cellulose. The relationship of structure to
function of these substances in animal cells and plant cells.

Biochemical tests using Benedict's solution for reducing sugars and non-reducing sugars and
iodine/potassium iodide for starch.

Previous knowledge
Sketch the structure of glucose ( & ) and fructose
Explain the term condensation polymerisation
Explain the term isomer
Sketch the condensation reaction between monosaccharides
Explain the terms condensation and hydrolysis
Name the components of maltose, lactose and sucrose
Name the polysaccharide that makes up plant cell walls
Name the storage carbohydrate in plant cells and explain how you would test and recognise its
presence
Name the storage carbohydrate in animals
Explain why sugar is stored as these large polymers
Name the bond formed in condensation reactions between monosaccharides
Explain how you would test for glucose

Biofactsheets: 39, 159

Carbohydrates contain only the elements carbon, hydrogen and oxygen.

Monosaccharides
These all have the formula (CH2O)n, where n can be 3-7. The most common and
important monosaccharide is glucose, which is a six-carbon or hexose sugar, so has
the formula C6H12O6. Its structure is shown on the right.

The six carbon atoms are numbered as shown, so we can refer to individual carbon
atoms in the structure.

Disaccharides
Disaccharides are formed when two monosaccharides are joined together by a
glycosidic bond. The reaction involves the formation of a molecule of water (H2O).
This diagram shows two glucose molecules joining together to form the
disaccharide maltose. Because this bond is between carbon 1 of one molecule and
carbon 4 of the other molecule it is called a 1-4 glycosidic bond. Bonds between
other carbon atoms are possible, leading to different shapes, and branched chains.
There are three common disaccharides:. They are listed below with their
monomers shown
A closer look at polysaccharides: (starch, glycogen and cellulose). These are formed by the condensation polymerisation between many glucose units. We must
learn to describe their structure and how that structure is related to their function

Cellulose structure:

Is a polymer of glucose, which differs from glucose by the orientation of the hydroxyl
group (OH) at C1.
Condensation polymerisation occurs between the C1 and C4 of adjacent monomers
The monomers are held by1,4 glycosidic bonds.
Alternate beta glucose molecules are inverted
The chain is long and straight so can be closely packed with other chains
Adjacent chains can be held together by H-bonds

The -glycosidic bond cannot be broken by

amylase, but requires a specific cellulase
enzyme.
The only organisms that possess a cellulase
enzyme are bacteria, so herbivorous animals,
like
cows and termites whose diet is mainly
cellulose, have mutualistic bacteria in their guts
so that
The chain is long and straights can be packed close together they can digest cellulose. Humans cannot digest
Adjacent chains can be held together by H-bonds cellulose, and it is referred to as fibre.
This forms micro-fibrils which are rigid and can link to form cellulose fibres
Cellulose is a component of the cell wall
Its strength means it can resist osmotic pressure
Basic structure of starch
Storage polysaccharide
Insoluble (no effect on water potential and thus osmotically inactive)
Not a pure substance but a mixture of 2 polysaccharides
Amylose: a chain of alpha glucose held by 1,4 glycosidic bonds. It forms a helix, held
by H bonds within the chain

Amylopectin: a polymer of alpha glucose with 1,4 glycosidic bonds and a small
number of 1,6 branches. This gives it an open structure and the branches are quickly
hydrolysed

Starch: structure for function

Role = storage
Features: Insoluble stays in cell
Features: Osmotically inactive cell does not absorb water
Feature: good respiratory substrate provides many glucose molecules
on hydrolysis
Feature: Amylose is Coiled lots of glucose in a small space
Feature: amylopectin branched is rapidly hydrolysed to glucose
because enzymes can begin to operate on all brnaches
 Basic structure of glycogen

Glycogen is similar to amylopectin.

It is polymer of (1-4) alpha glucose with 9% (1-6) branches, though more than
starch. Because it is so highly branched, it can be mobilised (broken down by
glycogen phosphorylase to produce glucose for energy) very quickly, reflects the
grater metabolic demands of animal over plant

Animals storage polysaccharide

Found mainly in muscle and liver.

Starch/Glycogen vs Cellulose
Starch/Glycogen
1. (1,4 and) 1,6 bonds branches
2. All glucose same way up
3. Helix/coiled/compact
4. Alpha glucose
5. No (micro/macro) Fibrils/fibres

Celluose
1. 1,4 bonds no 1,6 bonds
2. Alternate glucoses upside down;
3. Straight;
4. Beta glucose;
5. Micro/macro fibrils/fibres;
All monosaccharides and most disaccharides (except sucrose) are reducing sugars.
To approximately 2 cm of test solution add an equal quantity of Benedicts reagent. Shake, and
heat for a few minutes at 95C in a water bath. A precipitate indicates reducing sugar. The
colour and density of the precipitate gives an indication of the amount of reducing sugar
present, so this test is semi-quantitative.

Describe how you could use Benedicts reagent to test a urine sample for the presence of
glucose.
Add blue Benedicts reagent (to sample) and heat;
if reducing sugar is present a colour change will be observed red/ brown/ orange/ green/
yellow;

Non reducing sugars are sugars that do not reduce benedicts solution, and must
be hydrolysed first. This is done with acid, hydrolyses the glycosidic bonds and
produces the monomers. The solution must then be neutralised as the benedicts
will not work in acidic conditions.

Describe a further biochemical test to find out if a substance is a non-reducing

sugar.
Heat with benedicts solution and get no change
Take a fresh sample and heat with acid (or hydrolyse using enzyme), then
neutralise (sodium hydrogen carbonate)
(heat) with Benedicts (solution);
Observe colour change
16. The activity of an enzyme can be measured by testing for the concentration of its product at regular intervals. Describe how the concentration
of a reducing sugar can be measured using a colorimeter.

1 using, standard / known, concentrations (of reducing sugar);

2 heat with, Benedicts (solution) / CuSO4 + NaOH;

3 (use of) same volumes of solutions (each time);

4 (use of) excess Benedicts;

5 changes to, green / yellow / orange / brown / (brick) red;

6 remove precipitate / obtain filtrate;

7 calibrate / zero, colorimeter;

A closer look at lipids
Specification

Triglycerides and phospholipids are two groups of lipid.

Triglycerides are formed by the condensation of one molecule of glycerol and three
molecules of fatty acid.

A condensation reaction between glycerol and a fatty acid (RCOOH) forms an ester bond.

The R-group of a fatty acid may be saturated or unsaturated.

In phospholipids, one of the fatty acids of a triglyceride is substituted by a phosphate-

containing group.

The different properties of triglycerides and phospholipids related to their different structures.

The emulsion test for lipids.

Students should be able to:

recognise, from diagrams, saturated and unsaturated fatty acids
explain the different properties of triglycerides and phospholipids.

Previous knowledge
Sketch the general structure of glycerol and fatty acid
Name the bonds formed in the condensation reaction between fatty acids and glycerol
Draw the condensation reactions between glycerol and fatty acids

Bio factsheets: 42, 152, 74

Sketch the condensation reaction between fatty acids and glycerol

 Lipids are a mixed group of hydrophobic compounds composed of the
elements carbon, hydrogen and oxygen.

Hydrophobic: water hating, non-polar molecules, lipid soluble

Hydrophilic: water loving, polar molecules, water soluble (not soluble in

lipids)

Triglycerides are commonly called fats or oils.

They are made of glycerol and 3 fatty acids.
Glycerol is a small, 3-carbon molecule with three alcohol groups. (C3H8O3). Glycerol is common to all
triglycerides and so the properties of the fats depends on the nature of the fatty acids.
Triglycerides are insoluble in water. They are used for storage, insulation and protection in fatty tissue (or
adipose tissue) found under the skin (sub-cutaneous) or surrounding organs. They yield more energy per unit
mass than other compounds so are good for energy storage.

Fatty Acids..

Have a carboxyl group and a hydrocarbon (CH2) chain that can be anything from C14-C22 units
long.

May have no C=C (double) bonds in the chain, making them saturated and straight chains

May have one (mono) or several (poly) C=C bonds, making them unsaturated. This causes kinks,
bends in the chains. This lowers the melting point. Plant and fish fats are liquids at room
temperature (oils) as they have a high degree of unsaturation. Animal fats are solids at room
temperature as they are saturated fats.
 Phospholipids
Similar o triglycerides, but a phosphate group in place of one fatty
acid chain.

The phosphate head is hydrophilic and polar (water soluble)

Two non-polar hydrophobic "tails" (the fatty acid chains).

They are the main components of the cell membranes. This dual
polar and no polar region makes them amphipathic/amphiphilic.

When mixed with water, phospholipids form droplet spheres with

the hydrophilic heads facing the water and the hydrophobic tails
facing each other. This is called a micelle.

Alternatively, they may form a double-layered phospholipid bilayer.

This traps a compartment of water in the middle separated from the
external water by the hydrophobic sphere. This naturally-occurring
structure is called a liposome, and is similar to a membrane
surrounding a cell.

Lipids (emulsion test). Lipids do not dissolve in water, but do

dissolve in ethanol. This characteristic is used in the emulsion test.
Do not start by dissolving the sample in water, but instead shake
some of the test sample with about 4 cm of ethanol. Decant the
liquid into a test tube of water, leaving any undissolved substances
behind. If there are lipids dissolved in the ethanol, they will
precipitate in the water, forming a cloudy white emulsion. The test
can be improved by adding the dye Sudan III, which stains lipids
red.
A closer look at proteins

Specification

Amino acids are the monomers from which proteins are made. The general
structure of an amino acid as:

Where NH2 represents an amine group, COOH represents a carboxyl group and R represents a
carbon-containing side chain. The twenty amino acids that are common in all organisms differ only in
their side group.

A condensation reaction between two amino acids forms a peptide bond.

Dipeptides are formed by the condensation of two amino acids.
Polypeptides are formed by the condensation of many amino acids.

A functional protein may contain one or more polypeptides.

The role of hydrogen bonds, ionic bonds and disulphide bridges in the structure of proteins.

Proteins have a variety of functions within all living organisms. The relationship between primary,
secondary, tertiary and quaternary structure, and protein function.

The biuret test for proteins.

Students should be able to relate the structure of proteins to properties of proteins named
throughout the specification.

Previous knowledge
Define a polymer
Explain what a condensation reaction is
Sketch the general formula for an amino acid
Sketch the condensation reaction between amino acids
Name the bond holding protein polymers together
Name the group of enzymes that digest proteins
Name the reagent used to test for proteins
Give some examples of types of proteins/roles of proteins
Explain where proteins are synthesised inside a cell
Explain why cells manufacturing proteins may have a large number of mitochondria

Bio factsheets: 80
 Proteins are made of amino acids. Amino acids are made of the five elements
CHONS

Amino acids are so-called because they have both amino groups and acid
groups, which have opposite charges. At neutral pH (found in most living
organisms), the groups are ionised as shown, so there is a positive charge at
one end of the molecule and a negative charge at the other end. The overall
net charge on the molecule is therefore zero. A molecule like this, with both
positive and negative charges is called a zwitterion.

There are 20 different R groups, and so 20 different amino acids. Since each R
group is slightly different, each amino acid has different properties, and this in
turn means that proteins can have a wide range of properties.

Some amino acids and their R groups are shown in the diagram on the right

There is also the general structure of an amino acid shown

 Polypeptides

Amino acids are joined together by peptide bonds. The reaction involves the formation of a molecule of water in another condensation polymerisation reaction. When
two amino acids join together a dipeptide is formed.

In a polypeptide there is always one end with a free amino (NH3) group, called the N-terminus, and one end with a free carboxyl (CO2) group, called the C-terminus.

Protein synthesis takes place in ribosomes. The sequence of amino acids in a polypeptide chain is determined by the sequence of the genetic code in DNA.

Sketch the condensation reaction between

two amino acids

Testing for proteins

 Protein Structure: Polypeptides are just a string of amino acids, but they fold up to form the complex and well-defined three-dimensional structure of working proteins. To
help to understand protein structure, it is broken down into four levels:

Tertiary structure
Primary structure Further folding of the secondary structure. The tertiary structure is
This is just the held together by bonds between the R groups of the amino acids in
sequence of amino the protein, and so depends on what the sequence of amino acids is.
acids in the There are three kinds of bonds involved:
polypeptide chain but Hydrogen bonds, which are weak.
this will determine the Ionic bonds between R-groups with positive or negative charges,
rest of the protein which are quite strong.
structure sulphur bridges - covalent S-S bonds between two cysteine amino
acids, which are strong.

So the secondary structure is due to backbone interactions and is thus

largely independent of primary sequence, while tertiary structure is
due to side chain interactions and thus depends on the amino acid
sequence. Thus mutations changing the primary structure may not
alter the secondary structure, could have a big impact on tertiary

Secondary structure
This is a result of protein folding, and consists of a few
basic motifs that are found in all proteins. The
secondary structure is held together by hydrogen bonds
between the carboxyl groups and the amino groups in
the polypeptide backbone. The two most common
secondary structure motifs are the -helix and the -
Quaternary structure
pleated sheet, but it does not have to be either of these
Association of several polypeptides such as
The -helix is held together by hydrogen bonds running
parallel with the long helical axis. There are so many Haemoglobin, antibodies, actin in muscle
hydrogen bonds that this is a very stable and strong
structure.
Because all amino acids have different R groups they have different solubilities and
different affinities for solids. For this reason they can be separated by CHROMATOGRAPHY.

A container is filled to a depth of 2 cm with a suitable chromatography solvent. The

container is sealed and left to allow the solvent to saturate the atmosphere. This is
important because solvent must not evaporate from the chromatogram as it is running.
A strip of chromatography paper has a pencil line drawn on it slightly more than 2 cm
from the bottom and small crosses are drawn on it to indicate where to add the amino
acids.
It is important not to touch the paper as there are amino acids on your fingers.
A spot of amino acid is applied to a cross on the line using a capillary tube, the spot must
not exceed 2mm in diameter.
The spot is dried with a hair drier and another is applied over the top. This is done until a
small but concentrated dot of amino acid has been built up on the cross.
The nature of the amino acid is indicated by writing in pencil below the spot.
This is repeated until each amino acid dot has been added.
The strip of chromatography paper is placed in the container, making sure that the solvent
doesnt splash above the line.
The solvent is drawn up the paper by capillary action but does not evaporate from it due
to the saturated air around it. The amino acid was be carried by the solvent and each will
travel at different speeds.
When the solvent has nearly reached the top the paper is removed and a line is drawn to
show where the solvent has reached, this is the SOLVENT FRONT.
The chromatogram is dried in a fumes cupboard and then sprayed with NINHYDRIN
SPRAY. When the chromatogram dries, the amino acids will appear as purple spots at
different distances up the chromatogram.
The distance each spot has traveled from the centre of the cross to the centre of the spot is
measured, call this D1.
The distance the solvent front has traveled from the cross is measured, call this S.
The Rf value can then be calculated as Rf = D1/ S
Each amino acid will have a unique Rf value and as the Rf values for all amino acids are
known they can be identified knowing the Rf value
 Quick test 1

1. Draw the general structure for a fatty acid (1)

2. Draw the general structure of glycerol (1)

3. Draw the general structure of an amino acid and label the amino group and carboxyl group
(1)

4. Draw alpha and beta glucose (1)

5. The process through which monomers combine to form a polymer (1)

6. The name of the bonds in (3)

a. A disaccharide
b. A polypeptide
c. Triglyceride

7. How does a phospholipid differ to a triglyceride? (1)

8. Describe a biochemical test to show the presence of a non-reducing sugar (3)

9. Describe a biochemical test to show the presence of a fat (2)

10. What do you understand the term, primary structure of a polypeptide (1)

11. What type of bonds a formed in the folding of the secondary structure of a protein (1)

12. What types of bonds may be present in the tertiary structure (2)

13. What word describes a change in the primary structure of polypeptide (1)
Y
14. What are the monomers present in

a. Sucrose
b. Lactose
c. Maltose

15. Define a polymer

16. Copy and Label the diagram of the phospholipid shown on the right (3)

17. Molecules can be represented in different ways. Figure 1 shows a model of a fatty acid.
It shows the different atoms that make up the molecule. Label parts A and B (2)

A..............................

B..............................
Protein quick test

1. What are the monomers of proteins?

2. What is a polypeptide?
3. Draw the general structure of an amino acid
4. What sort of reaction links amino acids together?
5. What is the name of the bond that forms between amino acids?
6. Describe the tertiary structure of a protein
7. Describe the biochemical test for a protein
8. Name two common shapes formed during development of the secondary structure in a
protein
9. Name the types of bonds present in the secondary structure

Carbohydrate quick test

10. Draw the structure for alpha glucose

11. What molecule is released in a condensation reaction between two monosaccharides?
12. What is the name of the bond between two monosaccharides?
13. Describe the test for a non-reducing sugar
14. Describe a test to show the presence of starch

Triglyceride quick test

15. What are the components of a triglyceride?

16. Draw the basic structure of a fatty acid
17. Explain the difference between saturated and unsaturated fatty acids
18. Describe how a phospholipid differs to a triglyceride
19. Draw a molecule of glycerol
20. Describe a biochemical test for fats
Enzymes

Specification

Each enzyme lowers the activation energy of the reaction it catalyses.

The induced-fit model of enzyme action.

The properties of an enzyme relate to the tertiary structure of its active site and its ability to
combine with complementary substrate(s) to form an enzyme-substrate complex.
The specificity of enzymes
The effects of the following factors on the rate of enzyme controlled reactions enzyme
concentration, substrate concentration, concentration of competitive and of non-competitive
inhibitors, pH and temperature.

Students should be able to:

appreciate how models of enzyme action have changed over time
appreciate that enzymes catalyse a wide range of intracellular and extracellular reactions that
determine structures and functions from cellular to whole-organism level.

Previous knowledge
What are enzymes and how do they work?
What enzymes are made from?
Where enzymes are synthesised
Examples of enzymes
Primary, secondary, tertiary, quaternary protein structure
Sketch amino acids general formula
How to test for proteins
Explain the effect of temperature on the rate of enzyme activity
Explain the lock and key hypothesis
Explain the term denaturing
Suggest some reasons for using enzymes in industry and some reasons for not using them in industry

Practical
Effect of a named variable on the rate of an enzyme controlled reaction

Bio factsheets: 31, 43, 222, 163,

Enzymes are biological catalysts. They increase the rate of reactions by a factor of Reaction mechanism
between 106 to 1012 times. Substrate (S) is converted to product (P) by the enzyme

Enzymes are proteins E + S ES complex EP complex E + P

The reaction takes place in a small part of the enzyme called the active site, while
the rest of the protein acts as "scaffolding
The amino acids around the active site attach to the substrate molecule and hold
it in position while the reaction takes place. This makes the enzyme specific for
one reaction only, as other molecules won't fit into the active site.

Many enzymes need cofactors (or coenzymes) to work properly. These can be
metal ions (such as Fe2+, Mg2+, Cu2+) or organic molecules (such as haem, Lock and key hypothesis
biotin, FAD, NAD or coenzyme A). Many of these are derived from dietary
This suggests that the active site has a rigid shape and that only a substrate with the
vitamins, which is why they are so important. The complete active enzyme with
its cofactor is called a holoenzyme, while just the protein part without its co - correct complimentary shape can bind to the active site. However, this has its
factor is called the apoenzyme. limitations

Limitations of the lock and key

Does not easily explain how activation energy is lowered

Does not easily explain the role of competitive inhibitors

Does not easily explain the role of non-competitive inhibitors

So the induced fit model is a better mechanism to explain enzyme

activity
 Advantages of the induced fit model

Can explain how the activation energy is lowered, the stretching and
distorting of bonds or causing the closer orientation of reactive groups

Explain how non-competitive inhibitors can bind to a region away

from the active site and change its shape so that substrate can no
longer bind to the active site

Explains how competitive inhibitors can bind to the active site or other
Induced fit hypothesis molecules with similar shapes to the substrate
This suggests a more flexible active site that can change shape

The active site doesn't really fit the substrate (or the product) at all, but instead fits a
sort of half-way house, called the transition state. When a substrate (or product)
molecule binds, the active site changes shape to fit itself around the molecule more
closely. For example if a bond in the substrate is to be broken, that bond might be
stretched by the enzyme, making it more likely to break. As the reactants bind, the
changing shape of the active site brings reactive groups closer together. Alternatively
the enzyme can make the local conditions inside the active site quite different from
those outside (such as pH, water concentration, charge), so that the reaction is more
likely to happen.

Enzymes lower the activation energy

The energy level that the reactant molecules must overcome

before a reaction can occur
Factors affecting the rate of enzyme activity Suggest some of these: temperature, pH, enzyme and substrate concentration, competitive and non-competitive
inhibitors

As temp + = enzyme and substrate have more K.E As enzyme concentration increases so will the rate:
and collide more frequently (more ES complexes as there are more enzyme substrate complexes
form) and with the energy to overcome activation forming.
energy barrier.
At very high concentrations of enzyme the rate
An optimum temperature will be reached where remains constant as substrate becomes the
the rate is at its maximum limiting factor

High temperatures cause denaturation, due to

the breaking of breaking of bonds holding the
tertiary structure together (H bonds/disulphide
bridges/ionic bonds)
Active site altered (changes shape) substrate
cannot bind, no enzyme substrate complexes form
Due to the high turnover of enzymes, and the fact
they are not destroyed in reactions they only need
to be present in low quantities in the cells.

Deviations from the optimum pH cause a decrease

As the substrate concentration increases, the rate
in enzyme activity.
increases because more substrate molecules can
collide with enzyme molecules, so more reactions
Small deviations change the charge at the active
will take place. At higher concentrations the
site and affect the binding of the substrate
enzyme molecules become saturated with
substrate, so there are few free enzyme molecules,
Larger deviations can cause the hydrogen and ionic
so adding more substrate doesn't make much
bonds holding the tertiary structure together to
difference.
change and the enzyme denatures, meaning
enzyme substrate complexes can no longer form
Enzyme inhibitors

Inhibitors prevent the binding of substrate to active site;

therefore fewer enzyme-substrate complexes formed, reducing
the rate of their reactions.

They are found naturally, but are also used artificially as drugs,
pesticides and research tools. There are two kinds of inhibitors.

Competitive:
2 Similarity of shape of inhibitor and substrate;
3 Inhibitor can enter/bind with active site (of enzyme

Non-competitive:
4 Affect/bind to enzyme other than at active site;
5 Distorts shape of active site;

With competitive inhibitors increasing the substrate

concentration can allow enzyme rate to approach a normal rate
as the substrate will outcompete the inhibitor, this is not the
same with a non-competitive inhibitor as it is not competing for
the active site.

End product inhibition/Allosteric effectors

The activity of some enzymes is controlled by certain molecules binding to a specific
regulatory (or allosteric) site on the enzyme, distinct from the active site. Different
molecules can inhibit or activate the enzyme, allowing sophisticated control of the rate.
Only a few enzymes can do this, and they are often at the start of a long biochemical
pathway. They are generally activated by the substrate of the pathway and inhibited by the
product of the pathway, thus only turning the pathway on when it is needed.
 The best measurement is the initial rate - that is the
initial slope of the time-course. The slope is
Measuring the rate of enzyme reactions calculated

You need a signal to measure. Typical signals include colour changes,

pH changes, mass changes, gas production, volume changes or
turbidity changes

Substrate depletion (starts high decreases as substrate is used up

and ES complexes are less likely

Production formation starts low, increases as rate is high plenty

of ES complexes, slows as substrate diminishes
Q. Explain the effects of enzyme concentration, substrate concentration and
competitive inhibitors on the rate of an enzyme-controlled reaction.

E enzyme concentration ~

1 reaction (rate) increases with increased enzyme; A high / low

2 more active sites available;

3 in excess substrate / as long as enough substrate (molecules

available to occupy active site);

4 (as reaction progresses) the rate will decrease as substrate,

used up / becomes limiting; R plateau E
(3 max)

S substrate concentration ~

1 reaction (rate) increases with increased substrate; A high / low

2 more, molecules available to enter active site / ESC formed;

A more successful collisions

3 reaches point where all active sites occupied;

4 no further increase in rate / reaches Vmax; A plateau / levels off

5 enzyme conc. becomes limiting / unless add more enzyme; S

(3 max)

C competitive inhibitor ~

1 inhibitor has similar shape to substrate;

2 can, fit / occupy, active site;

3 for short time / temporary / reversible;

4 prevents / blocks, substrate from entering active site;

5 rate determined by relative concentrations;

6 little inhibition / rate little reduced, if substrate conc. > inhibitor conc.; ora

7 ref to chance of, substrate / inhibitor, entering active site;

8 effects can be reversed by increasing substrate conc.; C

(5 max)

general points ~

10 drawing a suitable graph to illustrate point made with labelled axes;

11 ref to optimum (rate); 9 max

QWC ~ legible text with accurate punctuation, spelling and grammar 1

Calculating pH of a solution:

Q. A solution has a hydrogen ion concentration of 1.8 10-5 M. What is the pH of the solution?

A. pH and hydrogen ion concentration are related so one may be calculated from the other using the
formula above. log10 is log to the base 10 and [H+] = concentration of the hydrogen ions in moles
per litre.

Type the hydrogen ion concentration figure into the calculator. To do this type 1.8, then press the
EXP (or 10x) key, type 5 then press the change sign (+/) key (see Figure 1). Your display should
now show 1.8-05

Step 2
Find log10 of this hydrogen ion concentration by pressing the log key.
Your display should now show 4.7447275

Step 3
Press the change sign key (+/)
Your display should now show 4.744 727 5, which is the pH. Round this to an appropriate number of
decimal places, for example to 4.74

Question
The pH of a solution is 8.6. What is the concentration of hydrogen ions present?

A. 2.5-9

Questions
1 For each equation, rearrange to make A the subject.
a C = B A (1 mark)
b log10 A = B (1 mark)
c log3 A = C (1 mark)

2 Work out the pH of solutions with the following hydrogen ion concentrations.
a 3.6 10-3 moles per litre (1 mark)
b 1 10-7 mol dm-3 (1 mark)
c 10-12 M (1 mark)
d 0.06 mmol per litre (1 mark)
e 10-4mmol/L (1 mark)

3 Work out the hydrogen ion concentrations of solutions with the following pH values.
a 3.8 (1 mark)
b 9.6 (1 mark)
c 1.8 (1 mark)
d 11.34 (1 mark)
e 7.2 (1 mark)
Answers
1 a A = B C b A = 10B c A = 3-C (1 mark each)
2 a 2.44 b 7.0 c 11.0 d 4.2 e 6 (1 mark each)
3 a 1.58 10-4 (1 mark)
b 2.51 10-10 (1 mark)
c 0.016 (1 mark)
d 4.57 10-12 (1 mark)
e 6.3 10-8 (1 mark)
Questions
4 a Name the type of reaction that would occur when:
i monomers join to form a biological polymer (1 mark)
ii a biological polymer is separated into monomers. (1 mark)
b Two monosaccharides can join to form a disaccharide.
i Complete the word equation.
glucose fructose (2 marks)
ii Name the type of bond formed between glucose and fructose. (1 mark)
iii Describe how you would test for the presence of glucose in a food sample. (2 marks)
c i Describe the chemical difference between saturated and unsaturated fatty
acids. (2 marks)
ii Name one part of the cell where phospholipids are found. (1 mark)
5 The enzyme amylase catalyses the following reaction:
starch water maltose
a A student carries out this reaction and adds a few drops of iodine in potassium
iodide to the reaction mixture.
i State what colour would be seen if all the starch had been digested. (1 mark)
ii The student carried out the same reaction using amylase that had been
boiled beforehand. State, and explain, what colour would be seen after
adding iodine to this reaction mixture. (3 marks)

b Amylase has tertiary structure but no quaternary structure.

i State the number of polypeptide chains in amylase. (1 mark)
ii Covalent bonds can be found in amylase. Suggest two other types
of bond that could be present in amylase. (2 marks)
6 a The table below shows some features present in biological molecules.
Complete the table by using a tick () to show if a feature is present.
amino starch fatty
acid acid
can be saturated or unsaturated
contains glucose monomers
contains the chemical group
COOH
(3 marks)

b A student designs an investigation to determine the effect of substrate

concentration on the rate of an enzyme reaction.
They decide to carry out the investigation at 35C.
i State two other variables that must be kept constant during the
investigation. (2 marks)
The results of the investigation are shown in the sketch graph below. The
student repeats the same investigation, but in the presence
of a competitive inhibitor of the enzyme.
 ii Copy the graph and sketch another line to show the results in the presence
of the competitive inhibitor. (2 marks)
c Explain how the structure of phospholipids enables them to form cell
membranes. (3 marks)

Answers
1 a i condensation (1 mark)
ii hydrolysis (1 mark)
b i sucrose (1 mark)
water (1 mark)
ii glycosidic (do not accept glucosidic) (1 mark)
iii Benedicts (reagent / solution) (1 mark)
turns (brick) red on heating (1 mark)
c i saturated: have no carboncarbon double bonds (1 mark)
unsaturated: have one or more carboncarbon double bonds (1 mark)
ii any correctly named membrane (1 mark)

2 a i orange / yellow (1 mark)

ii blue / black (1 mark)
because the enzyme had been denatured (1 mark)
so the starch is still present / starch was not digested (1 mark)
b i one (1 mark)
ii hydrogen (bonds) (1 mark)
ionic (bonds) (1 mark)
disulfide (bonds) (1 mark)
maximum 2 marks

3 a
amino starch fatty
acid acid
can be saturated or (1 mark)
unsaturated
contains glucose (1 mark)
monomers
contains the chemical (1 mark)
group COOH
b i enzyme concentration (1 mark)
pH (1 mark)
ii line starting at origin and having more gradual slope (1 mark)
 line meeting the plateau of uninhibited graph (1 mark)
c hydrophilic heads (1 mark)
hydrophobic tails (1 mark)
form a bilayer in water (1 mark)

Enzyme quick test

1. Explain in terms of activation energy why enzymes enable reactions to occur at lower
temperatures (2)
2. What is the main difference between the induced fit and lock and key model of enzyme
theory. Explain why the induced fit is a better model to enzyme activity (3)
3. What determines the shape of an enzyme active site? (2)
4. Why will an enzyme only bind with one substrate? (1)
5. If a mutation occurred in a gene that codes for an enzyme, explain why the enzyme may no
longer perform its function (2)
6. Explain why an increase in temperature increases the rate of enzyme activity (3)
7. Give two factors that may denature an enzyme (1)
8. What happens to an enzyme when it is denatured by heating, explain why it cannot catalyse
the reaction (3)
9. What does saturation point mean in an enzyme controlled reaction? (1)
10. Explain what happens to the rate of a reaction when the substrate concentration is
increased after the saturation point (2)
11. Explain the difference between competitive and non-competitive inhibitors (3)

Answers
1. Activation energy is lowered in the presence of an enzyme because, as the enzyme
starts to bind the substrate the active site changes shape to fit the substrate more
closely, this distorts the substrate molecule, stretching and stressing the bonds, making
it more likely to break them.
2. Lock and key suggests a rigid active site that is exactly complementary to the substrate,
in the induced fit model the active site is flexible and can change shape to fit the
substrate closely as it binds
3. The position of the bond formation during tertiary structure. Ultimately this is
determined by the primary sequence of the polypeptide
4. It can only bind substrates that are complementary to the active site
5. Alters the primary sequence and this may alter the positions where the bonds form
during development of the tertiary structure, altering the shape of the active site
6. More kinetic energy; more frequent and high energy collisions between enzyme and
substrate; more enzyme substrate complexes form; faster rate of reaction
7. High temperatures and extreme pH values
8. Bonds holding the tertiary structure together break (H-bonds), this alters the shape of
the active site, no longer complementary to the shape of substrate, no enzyme substrate
complexes form
9. All the active sites are occupied by substrate
10. Remains constant as the enzyme is limiting the rate of reaction all the active sites are
occupied
11. Non-competitive inhibitors bind a point on the enzyme other than the active site and as
such cause a change in the active site so it no longer complements the substrate,
preventing enzyme substrate complexes. A competitive inhibitor binds to the active site,
directly competing with the substrate, and when bound prevents enzyme substrate
complexes forming
Q9. Read the following passage. (enzymes and proteins structure)

Jobs Tears is a cereal plant which grows in the tropics. An unusual protein has been
found in its grains. This protein is unusual because it has two functions. It acts as both
an enzyme inhibitor and as an enzyme. As an inhibitor, the protein reduces the activity
of starch-digesting enzymes. The protein acts as an enzyme by breaking down chitin,
a polysaccharide found in
5 the walls of many fungi, to its monomers. Because of the resulting more negative water
potential in the cytoplasm of the fungus, this effectively leads to death by osmosis of
any fungus attacking the grain.

Our knowledge of the relationship between protein structure and function has led to
the development of the new technology of protein engineering. This involves changing
the amino
10 acid sequence of a protein and altering its tertiary structure. Altering the tertiary
structure changes the proteins properties. So far, we have been unable to produce a
protein with more than one function such as that found in Jobs Tears. We have had
success, though, in making some enzymes more stable and less prone to heat
denaturation. We have done this by substituting amino acids and allowing the
formation of additional chemical bonds.

Use information from the passage and your own knowledge to answer the following
questions.

(a) (i) The protein found in Jobs Tears breaks down chitin (line 4). What type of
chemical reaction is involved in breaking down chitin?

.............................................................................................................(1)

(ii) Breakdown of chitin leads to death by osmosis of fungi attacking the grain
(lines 6 - 7). Explain how.

.............................................................................................................

.............................................................................................................

.............................................................................................................

.............................................................................................................(2)

(iii) This protein does not break down the cell walls of the Jobs Tears plant.
Explain why.

.............................................................................................................

.............................................................................................................(1)

(b) Explain what is meant by the tertiary structure of a protein (line 10).

.............................................................................................................

.............................................................................................................(1)
(c) (i) Explain how heating an enzyme leads to it being denatured.

.............................................................................................................

.............................................................................................................

.............................................................................................................

.............................................................................................................(2)

(ii) How can protein engineering make enzymes more stable and less prone to
heat denaturation (line 13)?

.............................................................................................................

.............................................................................................................

.............................................................................................................

.............................................................................................................(2)

(d) Describe how the sequence of amino acids in part of the protein from Jobs Tears
could enable this protein to act as an enzyme inhibitor.

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................(6)
M9. (a) (i) Hydrolysis;
1

(ii) Water enters fungus (by osmosis);

Increases pressure inside fungus;
Cell wall no longer strong enough/present so cannot
withstand this;
max 2

(iii) Cell wall (of plant) not made of chitin/made of cellulose;

Enzyme is specific to chitin / will not break down cellulose;
1

(b) Way in which the whole protein/polypeptide is folded / shape adopted by

whole protein molecule / further folding of 2 structure;
Do not credit unqualified reference to three-
dimensional shape.
Reject third level /third sort.
1

(c) (i) More (kinetic) energy;

Bonds/specified bonds (holding tertiary structure) break;
2

(ii) Change amino acids;

Allowing formation of more hydrogen bonds/disulphide bridges;
2

(d) 1 Sequence of amino acids gives shape;

2 This is tertiary structure;
3 Has similar shape to substrate;
4 Fits / competes for active site;
5 Fits at site other than active site;
6 Distorting active site;
7 Therefore substrate will not fit (active site);
max 6
[15]
Q15. Read the following passage. (enzymes)

Aspirin is a very useful drug. One of its uses is to reduce fever and 5
inflammation. Aspirin does this by preventing cells from producing
substances called prostaglandins. Prostaglandins are produced by
an enzyme-controlled pathway. Aspirin works by inhibiting one of the
enzymes in this pathway. Aspirin attaches permanently to a
chemical group on one of the monomers that make up the active site
of this enzyme.

The enzyme that is involved in the pathway leading to the production 10

of prostaglandins is also involved in the pathway leading to the
production of thromboxane. This is a substance that promotes blood
clotting. A small daily dose of aspirin may reduce the risk of
myocardial infarction (heart attack).

Use information from the passage and your own knowledge to answer the
following questions.

(a) Name the monomers that make up the active site of the enzyme (lines 6
7).

........................................................................................................................(1)

(b) The diagram shows the pathways by which prostaglandins and

thromboxane
are formed.

(i) Aspirin only affects one of the enzymes in this pathway. Use information in
lines
5 7 to explain why aspirin does not affect the other enzymes.

...............................................................................................................

...............................................................................................................

...............................................................................................................

...............................................................................................................(2)
 (ii) Which enzyme, X, Y or Z, is inhibited by aspirin? Explain the evidence from
the passage that supports your answer.

Enzyme ................................................................................................

Explanation ...........................................................................................

...............................................................................................................

...............................................................................................................(2)

(c) Aspirin is an enzyme inhibitor. Explain how aspirin prevents substrate molecules
being converted to product molecules.

........................................................................................................................

........................................................................................................................

........................................................................................................................

........................................................................................................................

........................................................................................................................(2)

(d) Aspirin may reduce the risk of myocardial infarction (lines 8 12). Explain how.

...............................................................................................................

...............................................................................................................

...............................................................................................................

...............................................................................................................

...............................................................................................................

...............................................................................................................

.........................................................................................

...............................................................................................................

...............................................................................................................(3)
(Total 10 marks)
 M15. (a) Amino acid / amino acids ;
If anything else is given as well do not award mark.
1

(b) (i) 1. Affects one monomer / amino acid;

i.e. What is affected

2. Not found in all active sites;

i.e. Where it is found.
2. Must relate to active site. Enzyme is insufficient.
2

(ii) 1. X;

2. Enzyme in both pathways;

2 Award independently
2

(c) 1. Occupies / blocks / binds to active site;

i.e. What it does in terms of the active site.

2. Substrate will not fit / does not bind / no longer complementary to /

enzyme-substrate complex not formed;
1. Ignore references to change in shape and shape
of aspirin
molecule.
Ignore reference to competitive inhibitor i.e.
Consequence
required
2

(d) 1. Prevents / reduces formation of thromboxane;

1. Must prevent/reduce production.

2. Blood clots do not form / less likely to form;

2. Accept converse from this point onwards

3. (Do not block) coronary arteries / vessels;

4. Heart muscle / wall gets oxygen;

4. Reference to heart must be qualified.
3 max
[10]
DNA & RNA structure

Specification

Deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) are important information-carrying
molecules. In all living cells, DNA holds genetic information and RNA transfers genetic
information from DNA to the ribosomes.

Ribosomes are formed from RNA and

proteins.

Both DNA and RNA are polymers of

nucleotides. Each nucleotide is formed
from a pentose, a nitrogen-containing
organic base and a phosphate group:

The components of a DNA nucleotide are deoxyribose, a phosphate group and one
of the organic bases adenine, cytosine, guanine or thymine.
The components of an RNA nucleotide are ribose, a phosphate group and one of the
organic bases adenine, cytosine, guanine or uracil.
A condensation reaction between two nucleotides forms a phosphodiester bond.

A DNA molecule is a double helix with two polynucleotide chains held together by hydrogen
bonds between specific complementary base pairs.

An RNA molecule is a relatively short polynucleotide chain.

Students should be able to appreciate that the relative simplicity of DNA led many scientists
to doubt that it carried the genetic code.

Previous knowledge
Define a polymer
Explain what a condensation reaction is
Explain how DNA codes for proteins
Explain some basic structural features of DNA
Know what ribosomes are
The complimentary base pairs in DNA

Bio factsheets: 23, 115, 207

 The basic building block of DNA and RNA polymers are nucleotides.
The basic structure of a nucleotide is

A 5C pentose sugar (ribose = RNA/deoxyribose = DNA)

A phosphate group

A nitrogenous base
o In DNA bases are Adenine, Thymine, Cytosine & Guanine: A pairs with T and C pairs with G
o In RNA the bases are Adenine, Uracil, Cytosine, Guanine: A pairs with U and C pairs with G

Bases can be categorised as purines (double ring structures) or pyrimidines (single ring structures)
a purine always pairs with a pyrimidine. Thus the distance between each sugar phosphate
backbone in DNA is 3 rings. The bases are shown below
DNA/RNA polymerisation

Both DNA and RNA are polymers

Condensation polymerisation occurs between the nucleotides forming strong, covalent,

phosphodiester bonds. This results in a strong sugar phosphate backbone in the DNA
making it a stable molecule

DNA structure

Specific/complimentary base pairing occurs in DNA formation where

A pairs with T held by 2 H-bonds

C pairs with G held by 3 H-bonds

This complimentary base pairing ensures that replication of DNA is accurate (it will
also ensure accurate transcription of the genes for protein synthesis)

Individually these H bonds are weak and allow the DNA to be separated so
replication and transcription can occur

Collectively, the large number of H-bonds means that the DNA is a stable molecule

DNA is double stranded. This is important as each strand can act as a template in
replication, a semi conservative process, meaning that the DNA is copied
accurately

The DNA strands are antiparallel. On strand runs from the 5 prime (5) to the 3
and the other form the 3 to the 5.the 3 and the 5 are a reference to the Carbons
at the end of the DNA strand. DNA replication occurs from the 5 to the 3
direction

DNA structure was discovered by Watson and Crick

DNA replication Initially they were unsure if replication was conservative, dispersive or semiconservative

Conservative: means that the original strand remains as it was and a completely new strand is
synthesised that is identical

Dispersive replication: original DNA strands are dispersed/spread throughout the daughter DNA strands.
So the new DNA strands are a mixture of old and original strands

Semi conservative: Each strand acts as a template and the daughter DNA has one original strand and one
new strand

This was uncovered thanks to the work of Meselson & Stahl. (shown on the next page

DNA replication is semi conservative

This means that each strand acts as template in replication 9ensuring accuracy) and the resulting daughter
strands have one new and one original strand

The process
DNA helicase breaks the H-bonds between base pairs and DNA unzips
Free nucleotides align to their complimentary base pairs (AT & CG)
DNA polymerase joins the free nucleotides
Question

43. During research into the mechanism of DNA replication, bacteria were grown on
a medium containing nitrogen isotopes. The nitrogen isotopes used were heavy
nitrogen, 15N, and light nitrogen, 14N. After growth, the bacterial DNA was
isolated from the cells and spun in a centrifuge. The DNA settled in the
centrifuge tube at a position that corresponded to its density, indicating the
proportion of the different types of DNA present in the sample.

Bacteria were grown for many generations in a medium containing only the
heavy isotope of nitrogen, 15N. This resulted in all the DNA molecules
containing only 15N. The result after centrifugation is shown in Fig. 1.
14
These bacteria were then grown in a medium containing only light nitrogen, N.
After allowing time for the DNA to replicate once, the DNA was analysed as
before. The result is shown in Fig. 2.

centrifuge
tube 14 14
N DNA N DNA
14
N/ 15 N DNA 14
N/ 15 N DNA
15 15
N DNA N DNA

key: DNA

Fig. 1 Fig. 2

(a) Explain how this information supports the semi-conservative hypothesis of

DNA replication. [4]

The bacteria were allowed to continue to grow in the light nitrogen, 14N, until the
DNA had replicated once more. The DNA was analysed as before and the result
is shown in Fig. 3.

14
N DNA
14
N/ 15 N DNA
15
N DNA

Fig. 3
Fig. 4 shows simple diagrams of DNA molecules, indicating the nitrogen content
of each.

Key:
represents
DNA with 15N

A B C

represents
DNA with 14N

D E F

Fig. 4

(b) Select the letter or letters from Fig. 4 representing the bacterial DNA in Fig.
1, Fig. 2 and Fig. 3.

Fig. 1

Fig. 2.

Fig. 3...
[3]

(c) The bacteria were allowed to continue to grow in the light nitrogen, 14N,
until the DNA had replicated once more. The DNA molecules were
analysed as before. Complete the diagram to indicate the expected results
showing the composition of these DNA molecules.

14
N DNA

14 15
N N DNA

15
N DNA

[2]
[Total 9 marks]
Answer

43. (a) R first reference to 15N being radioactive

semi-conservative replication would give

1 one, template / original / old / parent, strand and one,

new / daughter, strand;

2 complementary base pairing / joining of new nucleotides /

other detail of forming the new strand;

data shows that

3 two isotopes in molecule / molecule contains both 14N and 15N;

4 one strand with, heavy N / 15N; R molecule

5 one strand with, light N / 14N; R molecule

6 no molecules with only, 1 isotope / 14N / 15N;

some points, particularly 4 and 5, could be awarded for a

correctly labelled or keyed diagram 4 max

(b) correct answer only - do not accept from a selection

A;
C;
C and E; 3

(c) 1 band = 0
3 bands = 0

band drawn for 14N and 14N/15N only;

thick for 14N and thin for 14N/15N; 2
[9]
Questions

1 Convert these fractions to decimals. Give your answer to 2 significant

figures.
3 16
a (1 mark) (1 mark)
12 56
2 What is the fraction of A in the following ratios?
a 5A : 10B (1 mark)
b 11B : 2A (1 mark)
3 What is the percentage of A in parts a and b of question 2? (2 marks)
4 In a sample of DNA, there was 17% thymine.
a Calculate the percentage of cytosine in the molecule. (1 mark)
b What is the percentage of purine bases in the sample? (1 mark)
5 A sample of DNA from the lambda bacteriophage virus was
sequenced and was found to be made up from 48502 base pairs.
12336 base pairs were adenine and thymine.
a How many of the bases in the molecule will be cytosine? (1 mark)
b Calculate the percentage of the DNA molecule which is made up
from guanine. (1 mark)
6 Pyrimidines make up 29.50% of a DNA sample.
a Find the percentage of adenine. (1 mark)
b Predict the percentage of guanine that will be found in a molecule
of mRNA that is made by transcribing this DNA sample. Assume
that all the bases are equally distributed on each of the two
strands in the DNA molecule. (1 mark)
7 A sample of messenger RNA was analysed and gave the following
base sequence:
AAGCGGUUACGGAAUAG
a What is the percentage of pyrimidines in the original coding DNA
strand? (1 mark)
b What proportion of bases in the original coding DNA molecule was
contributed by guanine? (1 mark)

Answers
1 a 0.25 (1 mark)
b 0.29 (1 mark)
2 a 5/15 (1 mark)
b 2/13 (1 mark)
3 A is 33.3% and 15.4% (2 marks)
4 a 33% (1 mark)
b 50% (1 mark)
5 a 36,166 (1 mark)
b 37.3% (1 mark)
6 a 35.25% (keep same number of decimal places as in the question) (1 mark)
b 7.38% (1 mark)
7 a 70.6% (12 of the bases in the coding strand are C + T) (1 mark)
b 23.5% (8 of the 34 bases in the two strands of the DNA molecule
were G) (1 mark)
Questions
1 a Name:
i the pentose found in a DNA nucleotide (1 mark)
ii the four bases found in DNA. (1 mark)
b Complete the sentence:
Three out of the four bases found in RNA are the same as those
in DNA.
The base which is different is in RNA and .. in
DNA. (2 marks)
c Name the type of bond formed between:
i the phosphate and pentose in a nucleic acid (1 mark)
ii the complementary bases in DNA. (1 mark)

2 a Put ticks () in the boxes to show where DNA and RNA can be
found in a eukaryotic cell.
nucleus cytoplasm

DNA

RNA
(2 marks)

b Copy and complete the table to show differences between the

structures of DNA and RNA.
DNA RNA

(3 marks)
c Describe the process of DNA replication. (5 marks)

3 a Complete the table using ticks () in the boxes to show whether

each statement applies to DNA, RNA, or both.
statement DNA RNA
hydrogen bonds hold the structure together
formed from nucleotides
contains ribose
(3 marks)
b RNA is made from a DNA template.
Write down the RNA sequence that would be made from this
sequence of DNA:

CCGTAGTAC
(1 mark)
 c The table shows the ratio of bases in the DNA from two sources.
One is from a virus and the other is from an animal.
source of DNA ratio of C : G ratio of A : T
animal 1:1 1:1
virus 0.72 : 1 1.20 : 1
i Explain the ratios from the animal DNA. (2 marks)
ii Suggest why the ratios from the virus DNA are different. (1 mark)

4 a Name two enzymes involved in DNA replication. (2 marks)

b The DNA from a species of grasshopper was found to contain
29% adenine.
Calculate the percentage composition of the other three bases. (2 marks)
c Describe how the structure of DNA enables it to replicate semi-
conservatively. (3 marks)

Answers
1 a ideoxyribose (1 mark)
ii
adenine
cytosine
guanine
thymine (1 mark)
b uracil (1 mark)
thymine (1 mark)
c i covalent (bond) (1 mark)
ii hydrogen (bond) (1 mark)

2 a nucleus cytoplasm

DNA (1 mark)

RNA (1 mark)

b
DNA RNA
contains thymine contains uracil (1 mark)
contains deoxyribose contains ribose (1 mark)
double stranded single stranded (1 mark)
c DNA helicase unwinds the double helix (1 mark)
hydrogen bonds broken (1 mark)
each strand acts as a template (1 mark)
new / free, nucleotides form complementary base pairs (1 mark)
(base pairs form) A with T and G with C (1 mark)
DNA polymerase forms covalent bonds between pentose and
phosphate (1 mark)
process continues for the entire molecule (1 mark)
 maximum 5 marks

3 a
statement DNA RNA
hydrogen bonds hold the (1 mark)
structure together
formed from nucleotides (1 mark)
contains ribose (1 mark)
b GGCAUCAUG (1 mark)
c i C G and A T are complementary (1 mark)
quantities of complementary bases must be equal (1 mark)
ii not double stranded / is single stranded / does not form base
pairs (1 mark)

4 a DNA helicase (1 mark)

DNA polymerase (1 mark)
b thymine / T 29% (1 mark)
guanine / G 21% and cytosine / C 21% (1 mark)
c double stranded / anti-parallel strands (1 mark)
weak / easily broken, hydrogen bonds (1 mark)
complementary base pairs / A-T and G-C (1 mark)
Genetics quick test 1
1. What do the terms locus/loci refer to on chromosomes
2. What is a homologous pair of chromosomes?
3. What is an allele?
4. What are introns?
5. What are exons?
6. What does diploid mean
7. What does haploid mean
8. What is the function of the histone proteins in DNA?
9. How are complimentary base pairs held together?
10. What are the base pairs?
11. What is the difference between a purine and pyrimidine base
12. How many codons are there?
13. What do these codons mean?
14. How many amino acids are there?
15. What is a codon?
16. How does eukaryotic and prokaryotic DNA differ?
17. What is a gene?
18. What is a mutation?
19. How can a mutation result in a non-functioning enzyme?
20. What are possible types of mutations?
21. What are mutagens and give some examples?
22. What does it mean to say the genetic code is degenerate?
23. What does it mean that the genetic code is non-overlapping?
ATP

Specification

A single molecule of adenosine triphosphate (ATP) is a

nucleotide derivative and is formed from a molecule of
ribose, a molecule of adenine and three phosphate
groups.

Hydrolysis of ATP to adenosine diphosphate (ADP) and an inorganic phosphate group (Pi) is catalysed
by the enzyme ATP hydrolase.

The hydrolysis of ATP can be coupled to energy-requiring reactions within cells.

The inorganic phosphate released during the hydrolysis of ATP can be used to phosphorylate other
compounds, often making them more reactive.

ATP is resynthesised by the condensation of ADP and Pi. This reaction is catalysed by the enzyme
ATP synthase during photosynthesis, or during respiration.

Previous knowledge
How is energy released in organisms?
The word and symbol equations for respiration and photosynthesis
Uses of energy is organisms
The concept hydrolysis and condensation reactions.

Bio factsheets: 129

ATP
A nucleotide derivative
The enzyme catalysing its hydrolysis is called ATP
hydrolase
The enzyme catalysing its production is ATP synthase

ATP synthesis

ATP is the energy currency of the cell. It is synthesised in respiration

in all organisms and can also be synthesised in photosynthesis in
plants (a process called photophosphorylation) during a stage of the
process called the light dependent reactions.
Possible uses of ATP in organisms
Active transport; Phagocytosis; Synthesis of glycogen (in liver and muscles); Protein / enzymes;
DNA / RNA; Lipid / cholesterol; Bile production (in liver); Cell division; exocytosis
Muscle contraction

ATP can be resynthesized in animals by the hydrolysis of a substance

called creating phosphate (CP)
Uses of ATP
CP C + P + energy

ADP + P (+ energy) ATP

This is a coupled reaction where the phosphate and energy from

stage 1 are used to resynthesize the ATP

ATP characteristics: It is described as energy intermediate or an

immediate energy source because.

It release energy rapidly in a single step hydrolysis

Directly to the reaction requiring it
In small manageable quantities
The phosphate can be added to other molecules, phosphorylation,
and this can activate them, making them more reactive
Water and inorganic ions

Specification

Water is a major component of cells. It has several properties that are important in biology. In
particular, water:
is a metabolite in many metabolic reactions, including condensation and hydrolysis reactions
is an important solvent in which metabolic reactions occur
has a relatively high heat capacity, buffering changes in temperature
has a relatively large latent heat of vaporisation, providing a cooling effect with little loss of
water through evaporation
Has strong cohesion between water molecules; this supports columns of water in the tube-
like transport cells of plants and produces surface tension where water meets air.

Specification

Inorganic ions occur in solution in the cytoplasm and body fluids of organisms, some in high
concentrations and others in very low concentrations.

Each type of ion has a specific role, depending on its properties.

Students should be able to recognise the role of ions in the following topics: hydrogen ions and pH;
iron ions as a component of haemoglobin; sodium ions in the co-transport of glucose and amino
acids; and phosphate ions as components of DNA and of ATP.

Bio factsheets: 30,

Water is an important constituent of the tissues. Typically about 60% of the water we take in comes
from drinks, 30% from food and the remaining 10% is metabolic water (a by-product of respiration).

Water molecules are charged, with the oxygen atom being slightly negative ( -) and the hydrogen
atoms being slightly positive ( +). These opposite charges attract each other, forming hydrogen
bonds that bind water molecules loosely together.

They are formed between an atom (usually hydrogen)

with a slight positive charge (denoted d+) and an
atom (usually oxygen or nitrogen) with a slight
negative charge (denoted d). Because hydrogen
bonds are weak they can break and form
spontaneously at the temperatures found in living
cells without needing enzymes.

Because it is charged, water is a very good solvent, and almost all the chemical reactions of life take
place in aqueous solution.
Charged or polar molecules such as salts, sugars, amino acids dissolve readily in water and so
are called hydrophilic ("water loving").
Uncharged or non-polar molecules such as lipids do not dissolve so well in water and are
called hydrophobic ("water hating").

Many important biological molecules ionise when they dissolve (e.g. acetic acid acetate- + H+), so
the names of the acid and ionised forms (acetic acid and acetate in this example) are often used
loosely and interchangeably, which can cause confusion. You will come across many examples of two
names referring to the same substance, e.g. phosphoric acid and phosphate, lactic acid and lactate,
citric acid and citrate, pyruvic acid and pyruvate, aspartic acid and aspartate, etc. The ionised form is
the one found in living cells.

Water molecules "stick together" due to their hydrogen bonds, so water has high cohesion. This
explains why long columns of water can be sucked up tall trees by transpiration without breaking. It
also explains surface tension, which allows small animals to walk on water.

Properties and uses of water. The list below shows some of its properties and uses.

Hydrogen bonds are formed between the oxygen of one water molecule and the hydrogen of
another. As a result of this water molecules have an attraction for each other known as cohesion.
Cohesion is responsible for surface tension which enables aquatic insects like pond skaters to walk
on a pond surface. It also aids capillarity, the way in which water moves through xylem in plants.

Water is a dipolar molecule, which means that the oxygen has a slight negative charge at one end of
the molecule, and each hydrogen a slight positive charge at the other end.. Other polar molecules
dissolve in water. The different charges on these molecules enable them to fit into waters hydrogen
bond structure. Ions in solution can be transported or can take part in reactions.
Water is used in photosynthesis, so it is responsible for the production of glucose. This in turn is used
in the synthesis of many chemicals.

Water helps in the temperature regulation of many organisms. It enables the cooling down of some
organisms. Owing to a high latent heat of vaporisation, large amounts of body heat are needed to
evaporate a small quantity of water. Organisms like humans cool down effectively but lose only a
small amount of water in doing so. A relatively high level of heat is needed to raise the temperature
of water by a small amount due to its high specific heat capacity. This enables organisms to control
their body temperature more effectively.

Water is a solvent for ionic compounds. A number of the essential elements required by organisms
are obtained in ionic form, e.g.: (a) plants absorb nitrate ions (NO3) and phosphate ions (PO4) in
solution (b) animals intake sodium ions (Na+) and chloride ions (Cl ).

A Water molecule consists of two Hydrogen atoms covalently bonded to an Oxygen atom.
Because oxygen is more electronegative than hydrogen, it has a greater pull on the shared
electrons. This that the oxygen atom is slightly negative (-) (because of the closer electrons),
and hydrogen is slightly positive (+). Water is therefore called a Polar Molecule.

The slightly negative and slightly positive regions of the water molecule are attracted to
charged regions of other molecules, forming Hydrogen Bonds (which are weak in comparison with
other chemical bonds). Water will form Hydrogen Bonds within itself.

Hydrogen bonds within water give it a high stability, which means that a large amount of
energy is required to raise the temperature of water (related to specific heat capacity). This
property means that oceans and lakes provide a stable environment in which organisms can
live. This also means that a large amount of heat is required to evaporate water (latent heat of
vaporisation), so it is very useful in cooling,.

As water decreases in temperature, its molecules are less able to break the Hydrogen bonds, as
they have less kinetic energy. This means that a semi-crystalline structure is formed, which
holds the water molecules apart, making ice less dense than liquid water, such that it floats.
This means that it insulates the water beneath, allowing organisms in the liquid water to
survive.

Cohesion is the tendency of molecules within a substance to 'stick together'. Water has a high
Cohesion because of Hydrogen bonding. This is important as transport of water in the Xylem in
plants relies on water being pulled up. Cohesion also gives the water a high surface tension,
allowing small organisms, such as Pond Skaters, to walk along it.

Water is good solvent for other polar molecules since it can interact with the charged regions
and dissolve the substance. It is also a good solvent for ionic substances, since the water
molecules cluster around the ions and separate them, thus dissolving the substance. This
property, along with the fact that water is liquid over a wide range of temperatures, makes it
ideal for acting as a medium in which metabolic reactions can occur, and also as a transport
vehicle.

Water also takes part in some metabolic reactions, for example, in Hydrolysis and
Condensation reactions.
Water as a solvent
Water is an excellent solvent for ions and polar molecules (molecules with an uneven charge)
because the water molecules are attracted to them, collect around them and separate them, so that
they dissolve. The chemicals are then free to move around and react with other chemicals and most
processes taking place in living organisms, happen like this in solution. Non-polar molecules like
lipids do not dissolve in water and tend to be pushed together by it. This is important in hydrophobic
interactions in protein structure and in membrane structure. Due to it's ability to dissolve so many
molecules, water is an important transport medium in animals and plants.

Thermal properties
Water molecules are attracted to one another by hydrogen bonds and this restricts the movement
of the molecules. This means that a relatively large amount of energy is required to increase the
temperature of water (it has a high specific heat capacity) and that large bodies of water are slow to
change temperature e.g. lakes and oceans. Due to their high water content, the bodies of organisms
are also slow to change temperature and this makes maintaining a stable body temperature easier.
Water also requires a relatively high amount of energy to become a gas and this can be used as an
effective means of cooling the body by sweating and panting. Conversely, a relatively large amount
of energy must be transferred away from water to make it freeze, which is important for organisms
with high body water content and for those living in water.

Density and freezing properties

Water is unusual because it's solid form is less dense than it's liquid form. Below 4c the density of
water starts to decrease and so ice floats on water and insulates the water below it. This reduces the
chances of large bodies of water completely freezing and increases the chances of life in water
surviving. These changes in density of water with temperature are important in the oceans because
they set up currents, which circulate nutrients.

High surface tension and cohesion

Water molecules tend to stick together and this is exploited in the way that water moves in long
unbroken columns through the xylem tissue of plants and is an important property in cells. The
cohesion of water molecules generates a surface tension at the surface of water enabling small
organisms e.g. pond skaters to exploit it as a habitat.

Water has a number of important properties essential for life. Many of the properties below are due
to the hydrogen bonds in water.

Specific heat capacity. Water has a specific heat capacity of 4.2 J g-1 C-1, which means that it
takes 4.2 joules of energy to heat 1 g of water by 1C. This is unusually high and it means
that water does not change temperature very easily. This minimizes fluctuations in
temperature inside cells, and it also means that sea temperature is remarkably constant.
Latent heat of evaporation. Water requires a lot of energy to change state from a liquid into
a gas, and this is made use of as a cooling mechanism in animals (sweating and panting) and
plants (transpiration). As water evaporates it extracts heat from around it, cooling the
organism.
Density. Water is unique in that the solid state (ice) is less dense that the liquid state, so ice
floats on water. As the air temperature cools, bodies of water freeze from the surface,
forming a layer of ice with liquid water underneath. This allows aquatic ecosystems to exist
even in sub-zero temperatures.
Cohesion. Water molecules "stick together" due to their hydrogen bonds, so water has high
cohesion. This explains why long columns of water can be sucked up tall trees by
transpiration without breaking. It also explains surface tension, which allows small animals
to walk on water.
Ionization. When many salts dissolve in water they ionize into discrete positive and negative
ions (e.g. NaCl Na+ + Cl-). Many important biological molecules are weak acids, which also
ionize in solution (e.g. acetic acid acetate- + H+). The names of the acid and ionized forms
 (acetic acid and acetate in this example) are often used loosely and interchangeably, which
can cause confusion. You will come across many examples of two names referring to the
same substance, e.g.: phosphoric acid and phosphate, lactic acid and lactate, citric acid and
citrate, pyruvic acid and pyruvate, aspartic acid and aspartate, etc. The ionized form is the
one found in living cells.
pH. Water itself is partly ionized (H2O H+ + OH- ), so it is a source of protons (H+ ions), and
indeed many biochemical reactions are sensitive to pH (-log[H+]). Pure water cannot buffer
changes in H+ concentration, so is not a buffer and can easily be any pH, but the cytoplasms
and tissue fluids of living organisms are usually well buffered at about neutral pH (pH 7-8).

1. liquid at normal temperatures ;

2. hydrogen bonding between water molecules ;
3. molecules more difficult to separate ;
4. ice floats on water / water freezes from top down ;
5. insulates water beneath ;
6. large bodies of water dont freeze completely / animals can still swim etc ;
7. (change in density with temperature) causes currents to circulate nutrients ;
8. solvent for , polar / ionic , substances ;
9. solubility of gases in environment ;
10. allows reactions to take place ;
11. transport medium ;
12. e.g. (of substance carried in what) ;
13. transport medium for , gametes / blood cells ;
14. water slow to change temperature ;
15. lakes / oceans / large volumes , provide thermally stable environment ;
16. internal body temperature changes minimised ;
17. used for cooling ;
18. e.g. (sweating / panting / transpiration) ;
19. large amount of energy must be removed for water to freeze ;
20. organisms can use surface of water (as habitat) ;
21. e.g. ; (of organism)
22. can form (long / unbroken) columns of water ;
23. ref. to vascular tissue / xylem ;
24. reactant (photosynthesis) ;
25. role in , hydrolysis / condensation ;
26. AVP ; e.g. transparency
27. AVP ; plants can photosynthesise under water
incompressible
hydrostatic skeleton / turgor
buoyancy
guard cell mechanism
support for large organisms on ice (penguins / polar bears)
further detail of any point
Water has a high specific heat capacity meaning it requires a lot of energy to raise its temperature
and it retains heat energy better than many substances. Thus its temperature does not fluctuate
rapidly and is a very important property for cold blooded animals. Also in aquatic systems water
does not heat up or cool down rapidly so lake and ocean temperature stays relatively constant.

Water requires a lot of energy to convert it form liquid to vapour. When it does evaporate it absorbs
large amounts of heat from the surroundings, hence its effectiveness in cooling bodies through
sweating.

Inorganic ions

POTASSIUM salts are very soluble and are important in the correct functioning of nerves and
muscles. They are osmotically active in that they reduce the water potential.
CALCIUM salts are insoluble and they are often structurally important. They are a
component of bone and teeth as a complex salt of phosphorous. Calcium pectate acts as a
glue sticking plant cell walls together. A flow of calcium ions is a component in the
functioning of the muscles.
MAGNESIUM is the metal ion in the porphyrin ring in chlorophyll is magnesium. It is a
constituent of the mineral component of bones and teeth
IRON is the metal ion in the porphyrin ring in haemoglobin is iron. It is involved in the
cytochromes involved in the electron transport chain in respiration.
HYDROGEN CARBONATE is involved in buffering systems in the blood.
NITRATES are essential nutrients, they are component in many biologically important
compounds such as amino acids which make up proteins and in the organic bases which are
in all nucleotides which in nucleic acids (DNA, RNA, NAD and ATP). Nitrogen is also in the
porphyrin rings of chlorophyll and haem molecules.
PHOSPHATES are a component in many biologically important compounds such as nucleic
acids, ATP, FAD, NAD and NADP. It is also a component of the complex calcium salt in bones
and teeth. Phospholipids are compounds of fat and phosphate and all cell membranes are
made of these.

Below 40C the density of water decreases and so it will
float. The molecules are less able to break the H-bonds
and a semi crystalline structure forms. As water cools,
more bonds form between the water molecules, each
water molecules can form 4 H- bonds with
neighbouring water molecules. To enable this bonding
the water molecules spread out and as such become
less dense. Ice floats on water. Water freezes from the
top down so Ice will act as an insulator for the
organisms in the water below allowing them to still
swim; it will allow gametes to move in external
fertilisation. The density changes in water will also set
up currents in the water circulating nutrients.
Cohesive (H-bonding) forces mean water is not
easily compressed. Useful in transpiration stream in
plants, also explains surface tension and how some
organisms can stand on water
Transparency: light can penetrate
to certain depths (depth depends
on wavelength) in aquatic
systems, thus photosynthesis can
occur.
Synovial fluid: in joints
Water interacts with the charged regions on polar
Pleural fluid: reduce friction between lungs and molecules, allowing them to dissolve into solution so
thoracic cage they can move around and react with other chemicals
in solution. Transport medium for polar molecules
Mucus (minerals into plants at the roots, through plants, in
the blood stream, removal of metabolic waste.
High degree of H-bonding in water gives it stability,
restricting the movement of the molecules. High
Water is used directly
specific heat capacity means large bodies of water are
In photosynthesis light dependent reactions relatively thermal stable, so body temperature remains
providing electrons for photosystems and for quite constant, as the temperature in cells remains
Hydrogen for reduction quite constant (more important in cold blooded)
aquatic environments stay more constant
In respiration, provides oxygen as terminal electron temperatures, enzyme activity Is temperature
acceptor in ATP synthesis sensitive, also solubility of gases will remain quite
constant. High latent heat of vaporisation, means that
Hydrolysis reactions like in digestion
body can be cooled considerably with only a minimal
loss of water
 H
H
Questions on water

O
1. The figure below represents a water molecule. Water molecules are
polar. As a result, they attract each other. Draw a second water molecule on the figure
above.

Your drawing should show:

the bond(s) between the two molecules

the name of the bond
the charges on each atom. [Total 3 marks]

2. Ponds provide a very stable environment for aquatic organisms. Three properties of
water that contribute to this stability are as follows:

the density of water decreases as the temperature falls below 4 C so ice floats on
the top of the pond
It acts as a solvent for ions such as nitrates (NO3)
A large quantity of energy is required to raise the temperature of water by 1 C.

Explain how these three properties help organisms survive in the pond. [Total 8 marks]

3. Water is important in many biological reactions.

Complete the table below by writing an appropriate term next to each description.

description term

the type of reaction that occurs when

water is added to break a bond in a
molecule

the phosphate group of a

phospholipid that readily attracts
water molecules

[Total 2 marks]
32. In this question, one mark is available for the quality of spelling, punctuation and
grammar.
The immense biodiversity of the oceans includes:
corals that host symbiotic algae which die if the sea temperature rises by 1C;
animals like polar bears that use floating ice as a base for sea fishing expeditions;
sessile animals like mussels that feed by filtering food particles from the water
and reproduce by releasing gametes into the water;
reef-building animals like corals that form hard calcium carbonate skeletons by
extracting mineral ions such as Ca2+ from the water;
seaweeds of different colours which occur in shallow water;
animals like fish that hunt prey using well-developed visual skills;
large animals such as the blue whale;
physically delicate organisms like jellyfish that lose their shape in air.

Using examples from the list above, describe and explain how the properties of water
make it a suitable environment for these organisms.
[8]

36. Complete the following passage by using the most suitable word(s) in each of the blank
spaces.

Water is essential for life. It makes up a high proportion of the cytoplasm in a cell. Many
different compounds can dissolve in it and it is therefore described as an excellent
..................................... . Water remains in the ....................... state over a wide range
of environmental temperatures. As it cools below 4 C it becomes less
............................. than warmer water. Ice floats on water, forming a layer that
................................... the water beneath with the result that large bodies of water
rarely freeze entirely. The ............................... bonds that form between water
molecules are responsible for its high .......................................... , which allows small
insects such as pond skaters to move on its surface without sinking.
[Total 6 marks]

56. Some of the properties of water are listed below.

boils at 100 C
freezes at 0 C
water below 4 C is less dense than water above 4 C
excellent solvent
much energy is required to raise the temperature of water
much energy is required to change water into water vapour
high surface tension and cohesion

Describe and explain the roles of water in living organisms and as an environment for
living organisms. You will gain credit for using information about the properties of
water. (Allow one and a half lined pages). [9]
Answers

.
+ hydrogen bond
+

H H

O -
O

-

H H

+

1 hydrogen bond represented as,

horizontal / vertical, dashed line between O on one molecule and H on the
adjacent molecule;
DO NOT CREDIT if >1 H bond is drawn between the
same two molecules

2 hydrogen / H, bond label (on any drawn bond between 2 molecules);

3 (delta positive) + on each drawn H

and (delta negative) (2) on each drawn O;
if both molecules drawn, + and on all atoms.
ACCEPT d (lower case) for
[3]

2. ice floats

P1 (ice less dense because) molecules spread out;

P2 molecules form, crystal structure / lattice / AW;

P3 ice forms insulating layer / clearly described; e.g. acts as a barrier to the cold

P4 water (below ice), does not freeze / still liquid / remains water / kept at higher
temperature;

S1 organisms do not freeze; DO NOT ACCEPT die (because survival stated in stem)

S2 animals / organisms, can still, swim / move;

S3 allows, currents / nutrients, to circulate;

solubility

P5 ions / named ion, polar / charged;

P6 ions /named ion, attracted to / bind to / interact with, water;

S4 (named) organisms / plants / animals, uptake / AW, minerals / named mineral /

nutrients; ACCEPT obtain / enters / goes in / gets

S5 correct use of named, mineral / nutrient, in organism;

 needs to be more specific than for growth /
metabolism suitable examples include but are not
limited to: nitrates for amino acids / protein / (named)
nucleic acid / phosphate for ATP / phospholipids /
plasma membrane / magnesium for chlorophyll etc

temperature stability

P7 many / stable, (hydrogen) bonds between molecules;

Many hydrogen bonds between molecules = 2 marks
(gets P7 and H)

P8 at lot of energy to, force apart molecules / break bonds;

ACCEPT heat as alternative to energy

P9 high (specific) heat capacity; DO NOT CREDIT latent heat capacity

S6 temperature does not change much / small variation in temperature;

could refer to organisms or surrounding water
ACCEPT stays cool in summer / stays warm in winter
DO NOT CREDIT constant alone

S7 effect of temperature on, enzymes / metabolic rate;

ACCEPT any reference to temperature affecting enzyme
activity / metabolic rate

S8 gases remain soluble;

Award once in any section

H hydrogen bonds;
DO NOT CREDIT if in incorrect context
(e.g. they are strong bonds)
7 max

QWC - Award if you see a P mark and an S mark within the same section;
Look for the S mark first, then award QWC if there is a P
mark in the same section in the mark scheme
1
[8]

3. hydrolysis / hydrolytic;
hydrophilic;
ACCEPT phonetic spelling throughout
IGNORE head
[2]
32. 1 hydrogen bonding;
2 detail; e.g. (electro)negative oxygen atom can hydrogen bond to
(electro)positive H atom/ one water molecule hydrogen bonds with
up to 4 others / H bonds individually weak / large collective effect
of many hydrogen bonds

coral algae
3 (high) thermal stability / temperature remains fairly constant;
4 water has high specific heat capacity;
5 much energy needed to break hydrogen bonds;

polar bears
6 cooling allows maximum number of hydrogen bonds to form;
7 water molecules space out to allow this;
8 water expands as it freezes / ice is less dense than water;

mussels, filter-feeders and sessile animals

9 water is transport medium for, food particles / gametes;
10 (tentacles / appendages / cilia) create currents bringing food;
11 ref. tides / ocean currents;
12 medium for, male gametes to swim / external fertilisation;
13 no desiccation of gametes;
14 ref to low viscosity / AW;

corals
15 minerals / ions, are soluble in water;
16 water is polar / detail of electrostatic attraction; A AW

seaweeds, fish eyes

17 water is transparent to light;
18 photosynthesis possible (in shallow water);
19 wavelength of light varies with depth;

whales, jellyfish
20 cohesion / water molecules stick to each other;
21 water not easily compressed;
22 gives support to large bodies / detail of upthrust or relative density;
23 acts as hydrostatic skeleton;
24 AVP; e.g. zonation / pigments
25 AVP; e.g. solubility of named gas linked to use in named organism 7 max

QWC legible text with accurate spelling, punctuation and grammar 1

[8]
36. solvent;
liquid; A same
dense;
insulates; A keeps warm R protects / warms
hydrogen; A H / weak R H+ / H2
surface tension / cohesion; 6

56. Marking points 1, 4, 8, 14, 19, 20 and 22 relate to the bullet points in the question

1 liquid at normal temperatures;

2 hydrogen bonding between water molecules;
3 molecules more difficult to separate;

4 ice floats on water / water freezes from top down;

5 insulates water beneath;
6 large bodies of water dont freeze completely / animals can still swim etc.;
7 (change in density with temperature) causes currents to circulate nutrients;

8 solvent for, polar / ionic, substances;

9 solubility of gases in environment;
10 allows reactions to take place;
11 transport medium;
12 e.g. (of substance carried in what);
13 transport medium for, gametes / blood cells;
Specification
The structure of eukaryotic cells, restricted to the structure and function of:
cell-surface membrane
nucleus (containing chromosomes, consisting of protein-bound, linear DNA, and one or
more nucleoli)
mitochondria
chloroplasts (in plants and algae)
Golgi apparatus and Golgi vesicles
lysosomes (a type of Golgi vesicle that releases lysozymes)
ribosomes
rough endoplasmic reticulum and smooth endoplasmic reticulum
cell wall (in plants, algae and fungi)
cell vacuole (in plants).

In complex multicellular organisms, eukaryotic cells become specialised for specific functions.
Specialised cells are organised into tissues, tissues into organs and organs into systems.

Students should be able to apply their knowledge of these features in explaining adaptations of
eukaryotic cells.

Previous knowledge
You should know the role of the
Mitochondria
Membrane
Cytoplasm
Nucleus
Ribosomes
Cell wall
Vacuole
Chloroplast
Plasmids (structures on bacterial cells)
Flagella (structures on bacterial cells)

You should know what the cell wall is made from

You should know the differences and similarities between plants, animal, bacterial and yeast cells
You should be able to understand the role of the different organelles and explain why different cells
have different quantities of these depending on the job of the cell (example: why pancreatic cells
may have a lot of ribosomes and mitochondria? Why muscle cells have a lot of mitochondria? Why
sperm cells have a lot of mitochondria located in a particular region)
You should be able to suggest how some cells are adapted to their function based upon their shape
and constitution
You should be able to define a tissue, organ, giving examples of these organs (stomach and leaf) and
the tissues within and their role.
You should understand that there is a hierarchy cells tissues organs systems organism
Explain what specialised cells are

Bio factsheets: 4, 61, 73, 107, 198,

Cells are the basic building block of life. Living organisms are classified into one of 5 kingdoms. The biggest division is between the cells of the prokaryote
kingdom (the bacteria) and those of the other four kingdoms (animals, plants, fungi and protoctista), which are all eukaryotic cells.

The biggest difference between a prokaryotic cell and eukaryotic cell is that prokaryotic cells do not have a membrane bound nucleus. Nor do they possess
membrane bound organelles (mitochondria, chloroplast etc)

Here are some general diagrammatic representations of

bacterial, plant and animal cells, and the some of the
structures (organelles) within them. We need to learn how the
structures outlined in the specification are adapted to do their
job.
 Cell Membrane (or Plasma Membrane). This is a thin, flexible layer round the outside of all cells made of
phospholipids (in a bilayer) and proteins (intrinsic and extrinsic).
It separates the contents of the cell from the outside environment, and controls the entry and exit of materials.

Lipid soluble (non-polar) molecules can move through the membrane by simple diffusion
Water soluble (polar molecules) move through the membrane using channel or carrier proteins. If this does not
use energy it is described as facilitated diffusion. Some carriers move molecules against the concentration
gradient and this requires ATP in a process called active transport.

Nucleus.
Largest organelle. Surrounded by a nuclear envelope, (double membrane)
Nuclear pores - large holes containing proteins that control the exit of substances such as RNA and ribosomes from
the nucleus.
The interior is called the nucleoplasm, which is full of chromatin (a DNA/protein complex in a 1:2 ratio) containing the
genes. During cell division the chromatin becomes condensed into discrete observable chromosomes.
The nucleolus is a dark region of chromatin, involved in making ribosomes.

Mitochondrion.
Rod-shaped organelle (8m long),
Site of aerobic respiration.
Mitochondria are surrounded by a double membrane: the outer membrane is simple and quite permeable, while
the inner membrane is highly folded into cristae, which give it a large surface area for reactions involved in ATP
synthesis to occur. The inner membrane is studded with stalked particles, which are the site of ATP synthesis.
The space enclosed by the inner membrane is called the mitochondrial matrix, and contains small circular strands
of DNA. The Mitochondria also has 70s ribosomes. This means that the mitochondria can synthesise the enzymes it
needs for reactions involved in aerobic repsiration
Chloroplast. Bigger and fatter than mitochondria, chloroplasts are where photosynthesis takes place.
They are enclosed by a double membrane, but chloroplasts also have a third membrane called the thylakoid
membrane. The thylakoid membrane is folded into thylakoid disks, which are then stacked into piles called
grana. The thylakoid membrane contains chlorophyll and other photosynthetic pigments arranged in
photosystems, together with stalked particles, and is the site of photosynthesis and ATP synthesis
The space between the inner membrane and the thylakoid is called the stroma.. Chloroplasts also contain
starch grains, ribosomes and circular DNA.
The chloroplasts contain chlorophyll and other accessory pigments that increases the range of wavelengths
that can be absorbed
It contains some genetic material and 70s ribosomes so it can synthesis enzymes needed for the process of
photosynthesis

Golgi Body (or Golgi Apparatus)

A series of fluid filled, flattened membrane sacs (Cisternae). Its job is to transport proteins from the RER and
sterols from the SER to the cell membrane for export.

Parts of the RER containing proteins fuse with one side of the Golgi body membranes, while at the other side
small vesicles bud off and move towards the cell membrane, where they fuse, releasing their contents by
exocytosis.

In the Golgi products from the SER and RER are modified and packaged. Modification may be the addition of
carbohydrates to form glycoproteins. It can also produce lysosome which contain hydrolytic (digestive
enzymes)

Lysosomes. These are small membrane-bound vesicles formed from the RER containing a cocktail of digestive
enzymes. They are used to break down unwanted chemicals, toxins, organelles or even whole cells, so that the
materials may be recycled. They can also fuse with a feeding vacuole to digest its contents.
Ribosomes. These are the smallest and most numerous of the cell organelles, and are the sites of protein synthesis. They
are composed of protein and RNA, and are manufactured in the nucleolus of the nucleus. Ribosomes are either found free
in the cytoplasm, where they make proteins for the cell's own use, or they are found attached to the rough endoplasmic
reticulum, where they make proteins for export from the cell. They are often found in groups called polysomes. All
eukaryotic ribosomes are of the larger, "80S", type.

Endoplasmic reticulum: system of interconnected flattened membranes. It can be smooth forming the
SER or studded with proteins forming the RER

Smooth Endoplasmic Reticulum (SER). Series of membrane channels involved in synthesising and
transporting materials, mainly lipids/sterols, needed by the cell.

Rough Endoplasmic Reticulum (RER). Similar to the SER, but studded with numerous ribosomes, which
give it its rough appearance. The ribosomes synthesise proteins, which are processed in the RER.

Cell Wall.
This is a thick layer outside the cell membrane used to give a cell strength and rigidity. Cell walls consist of a
network of fibres, which give strength but are freely permeable to solutes (unlike membranes).

Plant cell walls are made mainly of cellulose, but can also contain hemicellulose, pectin, lignin and other
polysaccharides. They are built up in three layers called the primary cell wall, the secondary cell wall and the
middle lamella.

There are often channels through plant cell walls called plasmodesmata, which link the cytoplasms of adjacent
cells.
Fungal cell walls are made of chitin (poly-glucosamine), bacterial cell walls are made of peptidoglycan. Animal
cells do not have a cell wall, though they do have a layer of carbohydrate outside the cell membrane called the
cell coat, or glycocalyx.
Vacuoles.
These are membrane-bound sacs containing water or dilute solutions of salts and other solutes.
Most cells can have small vacuoles that are formed as required, but plant cells usually have one very large
permanent vacuole that fills most of the cell, so that the cytoplasm (and everything else) forms a thin layer round
the outside.
Plant cell vacuoles are filled with cell sap, and are very important in keeping the cell rigid, or turgid. Some
unicellular protoctists have feeding vacuoles for digesting food, or contractile vacuoles for expelling water.
The membrane around the vacuole is called the tonoplast
Look back at the artist representations of the various organelles to try and compare against the pictures shown below generated using a special microscope called a
Transmission Electron Microscope
Specification
Prokaryotic cells are much smaller than eukaryotic cells. They also differ from eukaryotic cells in
having:
cytoplasm that lacks membrane-bound organelles
smaller ribosomes
no nucleus; instead they have a single circular DNA molecule that is free in the cytoplasm
and is not associated with proteins
a cell wall that contains murein/peptidoglycan, a glycoprotein.

In addition, many prokaryotic cells have:

one or more plasmids
a capsule surrounding the cell
one or more flagella.

Details of these structural differences are not required.

Viruses are acellular and non-living. The structure of virus particles to include genetic material,
capsid and attachment protein.

Previous knowledge

Eukaryotic cell structure and details of the organelles

A basic understanding of the bacterial cell (prokaryotic) structure

Biofactsheets: 32, 73, 181,

 Cytoplasm. Contains all the enzymes needed for all metabolic reactions, since there are no
organelles

Ribosomes. The smaller (70 S) type., protein synthesis

Nucleoid (or Nuclear Zone). The region of the cytoplasm that contains DNA. It is not
surrounded by a nuclear membrane.

DNA. Always circular, and not associated with any proteins to form chromatin, does not
contain introns (unlike eukaryotic DNA which has introns, is larger, is linear and is
associated with proteins).

Plasmid. Small circles of DNA, used to exchange DNA between bacterial cells, and very
useful for genetic engineering.

Cell membrane. made of phospholipids and proteins, like eukaryotic membranes.

Mesosome. A tightly-folded region of the cell membrane containing all the membrane-
bound proteins required for respiration and photosynthesis. Can also be associated with the
nucleoid.

Cell Wall. Made of murein (not cellulose), which is a glycoprotein (i.e. a

protein/carbohydrate complex, also called peptidoglycan). There are two kinds of cell wall,
which can be distinguished by a Gram stain: Gram positive bacteria have a thick cell wall
and stain purple, while Gram negative bacteria have a thin cell wall with an outer lipid layer
and stain pink.

Capsule (or Slime Layer). A thick polysaccharide layer outside of the cell wall, like the
glycocalyx of eukaryotes. Used for sticking cells together, as a food reserve, as protection
against desiccation and chemicals, and as protection against phagocytosis.

Flagellum. A rigid rotating helical-shaped tail used for propulsion. The motor is embedded
in the cell membrane and is driven by a H+ gradient across the membrane. Clockwise
rotation drives the cell forwards, while anticlockwise rotation causes a chaotic spin. This is
the only known example of a rotating motor in nature.

Bacteria can produce toxins that make us fell ill

Endotoxins (lipopolysaccharides) produced from the breakdown of bacteria (cell walls);
exotoxins (proteins) secreted (from living cells)
 F
Identify the structures labelled A to F and give one
function of each.
E

A ...............................................................

Function. ....................................................
D
A
..
B
B .............................................................

Function. ................................................
C
...

C ................................................................................................................................

Function. ....................................................................................................................

D ................................................................................................................................

Function. ....................................................................................................................

E .................................................................................................................................

Function. ....................................................................................................................

F .................................................................................................................................

Function. ....................................................................................................................
Viruses are the smallest and simplest of the microbes. They are acellular (not made of cells)
and, since they cannot reproduce (or do any of the seven characteristics of life) on their own,
they are considered non-living. They are in fact more like complex chemicals than simple
living organisms.

Viruses are obligate parasites that can only reproduce inside host cells which get damaged in
the process, leading to disease. A virus has been famously described as "a piece of bad news
wrapped up in protein". Viruses are thought to have arisen from lengths of DNA that became
separated from their cells.

A complete virus particle is called a virion. A virion is the dormant form of a virus that is transmitted
between cells. Virions are too small to see with a light microscope and were first seen in the 1930s
using the electron microscope. Once inside a host cell virions dismantle into their separate parts,
and the virus can be reproduced.

All virions contain:

Nucleic acid, which can be DNA or RNA, and single or double-stranded. Viruses are classified
according to the type of nucleic acid they contain. The nucleic acid typically codes for 5-100 proteins
(by comparison, the bacterium E. coli has about 4000 genes).

A protein coat called a capsid, made of subunits called capsomeres. If the capsid proteins are
closely bound to the nucleic acid, then the combination is called a nucleocapsid. Because capsids are
composed of many repeating subunits, they tend to have simple geometrical shapes, such as helix or
icosahedron (20 triangular faces).

Some virions have very simple structures containing nothing else, but many virions have
more complex structures, including:
Enzymes, required to replicate the viral nucleic acid or incorporate it into a host.
A lipid envelope, not made by the virus itself, but derived from a host cell membrane.
Matrix proteins to attach the capsid to the envelope.
Glycoproteins to allow the virus to attach to host cells.
Specification

Electron microscopes and scanning electron microscopes.

Measuring the size of an object viewed with an optical microscope.
The difference between magnification and resolution. Using the formula

Principles of cell fractionation and ultracentrifugation as used to separate cell components.

Students should be able to appreciate that there was a considerable period of time during
which the scientific community distinguished between artefacts and cell organelles.

Biofactsheets: 75,
There are thousands of different kinds of cell, but the biggest division is between the cells of the
prokaryote kingdom (the bacteria) and those of the other four kingdoms (animals, plants, fungi and
protoctista), which are all eukaryotic cells.
Prokaryotic cells are smaller and simpler than eukaryotic cells, and do not have a nucleus.
Prokaryote = without a nucleus
Eukaryote = with a nucleus

To see cells we need microscopes, like a light microscope. To see the ultrastructure of a cell (the
organelles inside) we need electron microscopes.

Converting between the various units of measurement in microscopy and using formula.

As units get smaller the numbers get bigger, for example 1cm 10mm 10,000m

Types of microscopy calculations

1. When given a scale bar: use the following formula

2. When you are not given a scale bar use the following formula

You must ensure your values are in the same units

You must be able to rearrange the formula
You must look for the units required in the answer and be able to convert between the
various units
Now try the following questions to practise using the different types of formula
 1. Figure 1 shows a section through a palisade cell in a
leaf as seen with a light microscope.
The palisade has been magnified 2000.

Figure 1

(a) Calculate the actual width of the cell, measured from A

to B, in m. Show your working

Answer ...........................................m
(2)

2. The drawing shows an electron micrograph of parts of epithelial cells from the
small intestine.

Mitochondria
0.1m

(b) (i) The scale bar on this drawing represents a length of 0.1m. Calculate the
magnification of the drawing. Show your working.

Magnification .............................................
(2)
3. (b) The drawing shows part of a plant cell as seen with an electron microscope.

(ii) Calculate the actual width of the cell from Y to Z. Give your answer in
micrometres (m) and show your working.

Answer ..................................... m
(2)

4. The diagram shows an animal cell.

20m
 (a) The scale bar on the diagram represents a length of 20 m. Use this to calculate
the magnification of this diagram. Show your working.

Magnification........................(2)

5. The diagram shows part of a plant cell as seen through an electron microscope.

(c) Calculate the width of the structure labelled X in micrometres.

Show your working.

Width ............................ micrometres (m)

(2)
(Total 7 marks)
6. The diagram shows part of an animal cell as seen through an electron
microscope.

(c) The actual diameter of organelle Y is 0.5 pm.m.

Calculate the magnification of the diagram. Show your working.

Magnification .....................................
(2)
(Total 7 marks)
7. (a) The drawing shows a section through a plant cell.

(ii) The magnification of the drawing is 7000. Calculate the maximum

actual length of the organelle labelled X. Show your working.

Answer ...................................... m
(2)

8. The diagram shows an Escherichia coli bacterium at a magnification of x 20 000.

Cytoplasm
A B

Capsule Cell wall Ribosomes Genetic material

(a) What is the actual length of the bacterium from A to B in micrometres (m)?

Show your working.

Length................micrometres (2)
9. The diagram shows an organelle from a palisade mesophyll cell, as seen with an
electron microscope.

Magnification: 38 000

(b) Calculate the maximum length of the organelle in micrometres. Show your
working.

Length: ............. micrometres (m) (2)

1. (a) 16 gains 2 marks; 2
(accept 15.5 . 16.5)
(principal of calculation i.e. measured distance
(31-33mm/3.1-3.3cm) gains 1 mark) Mag
2.

16 (1000) measuring scale bar

(b) (i) / principle of ; (15 17 tolerance)
0.1 dividing by 0.1

160000; (correct answer award 2 marks) 2

3.

(ii) 94/95/96 10; principle (measured distance Y-Z)

44/45/46 length of scale bar

20.4 21.8 2

4. (a) Two marks for correct answer of 3000;

One mark if answer obtained by dividing actual length by 20 m; 2

5. (c) 0.2 0.24 gains 2 marks;

ELSE evidence of observed measurement
(5 6 mm / 0.5 0.6 cm) 25 000
gains one mark; 2

6.

(c) 30 000 - 34 000 gains 2 marks;; 2

If neither of these: evidence of sensible measured diameter actual (0.0005 mm)

gains 1 mark

7.

(ii) Correct answer (3m = 2 marks)

error with measurement, but clearly derived by dividing drawing size
by magnification = 1 mark; 2

8. (a) 4 micrometres = 2 marks (Allow 3.9 - 4. 1)

Correct method of scaling, but incorrect conversion to micrometres = 1 2

9.

(b) 3.2 - 3.3 - gains 2 marks

Working shows measured length divided by magnification - gains 1 mark 2
 Types of microscope

Magnification and Resolving Power. By using more lenses microscopes can magnify by a larger
amount, but this doesn't always mean that more detail can be seen. The amount of detail depends
on the resolving power of a microscope, which is the smallest separation at which two separate
objects can be distinguished (or resolved). It is calculated by the formula:

Where is the wavelength of light, and n.a. is the numerical aperture of the lens (which ranges from
about 0.5 to 1.4). So the resolving power of a microscope is ultimately limited by the wavelength of
light (400-600nm for visible light). To improve the resolving power a shorter wavelength of light is
needed, and sometimes microscopes have blue filters for this purpose (because blue has the
shortest wavelength of visible light).

In essence:

There is no limit to how much you can enlarge an image, but as it gets larger often the image will become more
blurred (loses resolution). To magnify on a large scale and retain the detail you need to use a short wavelength and
electron microscopes allow this

Resolution: how close 2 points can be to each other and still be distinguished as 2 separate points.

Electron microscopes have a higher resolution than (light microscopes, as they use electrons that have a shorter wavelength
than light
Shorter wavelengths (like electrons) allow better resolution than longer wavelengths (like light).

See additional booklet for scaling questions involving the eye piece graticule and stage micrometer
 Light Microscope. This is the oldest, simplest and most widely-used form of
microscopy. Specimens are illuminated with light, which is focussed using glass
lenses and viewed using the eye or photographic film. Specimens can be living
or dead, but often need to be stained with a coloured dye to make them visible.
Many different stains for specific parts of the cell such as DNA, lipids,
cytoskeleton, etc. All light microscopes today are compound microscopes,
which means they use several lenses to obtain high magnification. Light
microscopy has a resolution of about 200 nm, (enough to see cells, but not the
details of cell organelles). There has been a recent resurgence in the use of light
microscopy, partly due to technical improvements, which have dramatically
improved the resolution far beyond the theoretical limit. For example
fluorescence microscopy has a resolution of about 10 nm, while interference
microscopy has a resolution of about 1 nm.

Electron Microscope. This uses a beam of electrons, to "illuminate" the

specimen produced using a hot wire, focussed (using electromagnets) and
detected (using a phosphor screen or photographic film). Short wavelength (< 1
nm), gives a high resolution allowing organelles such as mitochondria, ER and
membranes to be seen in detail
The main problem is that specimens must be fixed in plastic and viewed in a
vacuum, and must therefore be dead. Other problems are that the specimens
can be damaged by the electron beam and the stains (usually heavy metals like
osmium, lead or gold). Initially there was a problem of artefacts (i.e. observed
structures that were due to the preparation process and were not real), but
improvements in technique have eliminated most of these.
There are two kinds of electron microscope.
The transmission electron microscope (TEM): transmits a beam of electrons
through a thin specimen focussing the electrons to form an image on a screen or
on film. This is the most common form of electron microscope and has the best
resolution.
The scanning electron microscope (SEM): scans a fine beam of electron onto a
specimen and collects the electrons scattered by the surface. This has poorer
resolution, but gives excellent 3-dimentional images of surfaces.
Explain the advantages and limitations of using a transmission electron
microscope

Advantages
1 TEM uses (beam of) electrons;
2 These have short wavelength;
3 Allow high resolution/greater resolution/Allow more detail to be seen/greater
useful magnification;

Disadvantages
4 Electrons scattered (by molecules in air);
5 Vacuum established;
6 Cannot examine living cells;
7 Lots of preparation/procedures used in preparing specimens/ fixing/ staining/
sectioning;
8 May alter appearance/result in artefacts;
9 very thin specimens
10 black and white, images

How to use a microscope to measure the size of an object.

Measure with an eyepiece graticule

Calibrate with the stage mcirometer (an object of a known size)

Repeat and calculate an average

Cell fractionation: Separating different parts and organelles of a
cell, so that they can be studied in detail. The most common
method of fractionating cells is to use differential centrifugation.

Organelles are separated based on different densities

There are 3 key stages

1) Homogenisation
2) Filtration
3) Centrifugation
Describe how you would obtain a
sample of undamaged chloroplasts
Homogenisation: Pestle mortar (using and as abrasive component) from leaves.
In a blender
This breaks open cells. Done in a 1. Chop up leaves in a.
Buffered solution: Preventing pH changes that may damage the organelle 2. Cold: reduces enzyme activity
by denaturing enzymes or proteins affecting function 3. Buffered solution: prevents pH
changes affecting enzymes
Isotonic: prevent osmotic movement where swelling of organelle may 4. Isotonic: prevents osmosis and
cause lysis or shrinkage of organelle both resulting in destruction of possible lysis or shrinkage of organelles
natural function 5. Filter and centrifuge filtrate
6. Centrifuge supernatant
Cold (50C): This reduces enzyme activity and will prevent autolysis (self- 7. at higher speed;
destruction) of the organelle. 8. Chloroplasts in (second) pellet;

2) Filtration: removes any debris (unbroken cells, cartilage, and sand). This Key to remember you must separate
will prevent contamination of the pellets resulting from centrifugation organelles in order, most dense first
then least dense, if you try and jump to
a particular stage the sample will be
3) Centrifugation: Homogenate (solution of organelles) is centrifuged at contaminated
different speeds. Low speeds will cause more dense organelles to separate
and higher speeds will cause least dense organelles to sediment. Order in which organelles would sediment with most
dense to least dense
After spinning the pellet of organelle is at the bottom the remaining
solution is called the supernatant and poured (decanted) for Most dense organelle Nucleus
centrifugation. Chloroplasts/ Mitochondria
ER, Golgi, Lysosomes
Least dense organelle Ribosomes

80s in eukaryotic cells will sediment before 70s in prokaryotic

cells as they are larger (more dense)
Q14. Read the following passage. (investigating cells)

In a human, there are over 200 different types of cell clearly distinguishable
from each other. What is more, many of these types include a number of
different varieties. White blood cells, for example, include lymphocytes and
granulocytes.

Although different animal cells have many features in common, each type has
adaptations.
5 associated with its function in the organism. As an example, most cells contain
the same organelles, but the number may differ from one type of cell to
another. Muscle cells contain many mitochondria, while enzyme-secreting
cells from salivary glands have particularly large amounts of rough
endoplasmic reticulum.

The number of a particular kind of organelle may change during the life of
the cell. An
10 example of this change is provided by cells in the tail of a tadpole. As a
tadpole matures into a frog, its tail is gradually absorbed until it disappears
completely. Absorption is associated with an increase in the number of
lysosomes in the cells of the tail.

Use information from the passage and your own knowledge to answer the
following questions.

(a) Explain the link between.

(i) mitochondria and muscle cells (lines 6 - 7);

.............................................................................................................

.............................................................................................................

.............................................................................................................

.............................................................................................................(3)

(ii) rough endoplasmic reticulum and enzyme-secreting cells from salivary

glands (lines 7 - 8).

.............................................................................................................

.............................................................................................................

.............................................................................................................

.............................................................................................................(2)

(b) Use information in the passage to explain how a tadpoles tail is absorbed as a
tadpole changes into a frog.

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................(2)
 (c) Starting with some lettuce leaves, describe how you would obtain a sample of
undamaged chloroplasts. Use your knowledge of cell fractionation and
ultracentrifugation to answer this question.

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................

...................................................................................................................... (6)
(Total 13 marks)

M14. (a) (i) Mitochondria site of respiration;

Production of ATP / release of energy;
For contraction;
Do not award credit for making or producing
energy.
3

(ii) Enzymes are proteins;

Proteins synthesised/made on ribosomes;
2

(b) Lysosomes produce/contain enzymes;

Which break down/hydrolyse proteins/substances/cells of tail;
2

(c) 1. Chop up (accept any reference to crude breaking up);

2. Cold;
3. Buffer solution;
4. Isotonic / same water potential;
5. Filter and centrifuge filtrate;
6. Centrifuge supernatant;
7. At higher speed;
8. Chloroplasts in (second) pellet;
max 6
[13]
Specification

Within multicellular organisms, not all cells retain the ability to divide.

Eukaryotic cells that do retain the ability to divide show a cell cycle.
DNA replication occurs during the interphase of the cell cycle.
Mitosis is the part of the cell cycle in which a eukaryotic cell divides to produce two daughter cells,
each with the identical copies of DNA produced by the parent cell during DNA replication.

The behaviour of chromosomes during interphase, prophase, metaphase, anaphase and telophase of mitosis.
The role of spindle fibres attached to centromeres in the separation of chromatids.

Division of the cytoplasm (cytokinesis) usually occurs, producing two new cells.

Meiosis is covered in section 3.4.3

Students should be able to:

recognise the stages of the cell cycle: interphase, prophase, metaphase, anaphase and
telophase (including cytokinesis)
Explain the appearance of cells in each stage of mitosis.

Mitosis is a controlled process. Uncontrolled cell division can lead to the formation of tumours and
of cancers. Many cancer treatments are directed at controlling the rate of cell division.

Binary fission in prokaryotic cells involves:

replication of the circular DNA and of plasmids
Division of the cytoplasm to produce two daughter cells, each with a single copy of the
circular DNA and a variable number of copies of plasmids.

Being non-living, viruses do not undergo cell division. Following injection of their nucleic acid, the
infected host cell replicates the virus particles.

Previous knowledge

Principles of mitosis
Principles of meiosis
Compare meiosis and mitosis
Importance of mitosis
Explain the term mutation from a biological perspective
What are chromosomes made from?
Define a gene
Define an allele

Bio factsheets: 76, 237

The cell cycle
The life of a cell is called the cell cycle and has three phases:

Interphase (which can be further divided into)

o Growth 1 (G1)
o Synthesis (S)
o Growth 2 (G2)

Mitotic phase (which is subdivided into, prophase, metaphase, anaphase

and telophase

Cytokinesis

In interphase we must be able to explain or suggest some of the events happening

in each of the subdivisions

G1: cell increases in mass, new organelles and new proteins formed

S phase: DNA replication occurs (details of this will be covered in the course)

G2: ATP stores are increased, spindle proteins synthesised

Common questions asked

Approximately 90% of the cell cycle is interphase and the remaining 10% is
mitotic phase. Cell cycle times can vary. For example bacterial cells can divide
What happens in the S phase of interphase? DNA replication
every 30 minutes under suitable conditions, skin cells divide about every 12
Describe two events which occur during interphase. hours on average, liver cells every 2 years, and muscle cells never divide at all
Increased in volume of cell /cell bigger; after maturing, so remain in the growth phase for decades.
Increase in number of organelles;
Protein synthesis We can calculate how long the cell spends in each stage of mitosis because the
DNA replication number of cells at each stage is proportional to the time spent at that stage.
ATP synthesis / respiration;
The following formula may help
12/200*24 = 1.44hours

0.82*24 = 19.68 hours. 82% of the 24 hour cell

cycle was interphase

Short duration of interphase

Mitosis = 10% of the cell cycle,

10% of 20hours = 2 hours

Q. The time required for a cell to complete the cell cycle

was 4 hours 18 minutes. Calculate the time required in
minutes for this cell to multiply to produce eight cells.
Show your working.

A. 3 cycles thus 3*4 + 18*3 = 12 hours + 54 minutes

(74+18)/982 = 9.4%
 Prophase: Telophase:

chromosomes condense and become visible. Sister Chromosomes uncoil

chromatids are Held by centromere
New nuclear membrane forms around the
Nucleolus breaks down and nuclear membrane chromosomes
breaks down

There are early and late phases of this

Metaphase: (M = middle)

Chromosomes line up at the equator

Attach to the spindle fibre at the centromere

Cytokinesis:
Anaphase (A = apart)
Cellular division occurs and two genetically identical daughter cells
CHROMATIDS are separated to opposite poles are produced, containing a full set of chromosomes (diploid cells)
of cell

Spindle fibres contracts

Importance of mitosis
1. Growth / increase in cell number;
2. Replace cells / repair tissue / organs /body;
3. Genetically identical cells;
Microscope slides can be prepared to observe mitosis; this is done in the following way.
Use root tips (cell division is occurring here). Treat them with HCl (separation of the cells, kill 4. Asexual reproduction /cloning;
the cells and stop mitosis progressing). Squash the root tips (spreads the cells and makes them
thin enough for light to penetrate), stain with feulgen/acetic orcein (makes the chromosomes
visible)
 List in order (remember there are early and late stages of each phase)

Describe the role of the spindle in mitosis (2)

A D C E B;

Attachment of centromeres/chromosomes/chromatids; Separation of

centromeres/chromatids/chromosomes;

Name stages A and B. In each case describe what is happening to the chromosomes. (4)
Describe two events during interphase which prepare a cell for mitosis (2)

Prophase: chromosomes thickening/becoming visible;

Anaphase: chromatids moving to opposite poles/ ends of spindles;

DNA replication;
synthesis or proteins/build-up of energy stores/growth/increase in cytoplasm;
replication of organelles/named example;

List stages A to E in order starting with D

(D) B E A C;

Diagram shows cells in the stages of mitosis

Name stage A?

What evidence that B is in anaphase?

 Cancer treatment
Cell cycle and cancer development Aim is to disrupt parts of the cell cycle so cells will not divide and the tumour will not grow. The
Mutations are changes in the DNA, changes in the base sequence problem is that treatments harm healthy cells also. So treatments are given in short bursts to
coding for genes. This can be additions, deletions, substitutions. allow healthy cells to recover, but each time the tumour does not recover to the same extent.

They occur naturally but the frequency can increase with exposure to
mutagens (UV light, X-ray, gamma radiation, benzene)

If these mutations occur in genes controlling the cell cycle, the result
can lead to cancer.
Uncontrolled cell division leading to a tumour (mass of abnormal cells)

The cell cycle is controlled by two key genes

Proto-oncogenes: these code for proteins that encourage cell

divisions. If a mutation occurs in one of these, it forms oncogenes,
which means the proteins to encourage cell division are expressed at
all time, so cells are constantly trying to divide.

Tumour suppressor genes: these code for proteins that inhibit cell
division

Cancer requires mutations to arise in several genes involved in

regulating the cell cycle Approaches to treating cancer

Cancer cells are characterised by a high rate of division; they can Inhibit enzymes involved in DNA replication/prevent synthesis of enzymes involved in
separate from adjacent cells and migrate to other tissues rpelication: Helicase and polymerase, so that DNA replication is stopped, cell division is
stopped and reduces the spread of cancer

Inhibit nucleotide synthesis: stops DNA replication

Understanding the treatment of cancer requires and understanding of
the cell cycle and the role of certain components at each stage. Inhibit spindle formation/spindle shortening: stop chromosomes attaching at metaphase and
being separated

Radiation to damage DNA further: and when DNA is checking is done during interphase the
cell will kill itself
Calculating mitotic index

Introduction
When mitosis is actively occurring in a tissue, such as root tip meristems, it is
possible to make stained squashes to see the mitotic stages in individual cells. You
may do this as a practical using onion root tips. The mitotic index (MI) is a
proportion which is simple to find using the formula:

number of cells with condensed chromosomes

MI = 100
total number of cells
This will give you a percentage value.
Note that if the multiplication by 100 is not used, the index will be expressed as a
simple decimal, e.g. 24% would be expressed as 0.24.

Worked example
Question
In a stained sample of dividing cells, 29 cells were visible in one microscope field of
view. Three of these cells had clearly visible chromosomes. What is the MI of the
tissue?
Answer
Step 1
Substitute the counted numbers into the formula:
3
MI = 100
29
Step 2
Complete the calculation to find the MI:
(3 29) 100 = 10%

Questions
1 Calculate MI values for the following: (4 marks)
Number of cells with
Tissue sample Total number of cells
visible chromosomes
a 5 30
b 18 26
c 56 245
d 21 145
2 Calculate the MI of the tissue shown in the photograph on the
support sheet. (2 marks)
3 Mast Cell Tumours occur in dogs, often causing visible swellings on
the skin. The MI of samples taken from these tumours is helpful in
predicting likely survival times.
Predicted survival
MI (%)
times (months)
04 >70
048 <2
Biopsy tissue samples taken from tumours on two dogs were
examined and microscope cell counts were as follows:
Total number of Total number of cells
Dog Biopsy sample
cells counted in mitotic stages
1 492 369
Boxer 2 387 173
3 426 245
1 478 25
Labrador 2 429 22
3 351 11
a Calculate the MI for each sample. (6 marks)
b Calculate the mean MI for each dog. Show your working. (2 marks)
c Suggest a survival time for the boxer. (1 mark)
d Comment on the likely survival of the labrador. (2 marks)
Answers
1 a 16.7% b 69.2% c 22.9% d 14.5% (1 mark for each)
2 (10 42) 100 = 42 (counting whole cells only) (2 marks, 1 for calculation;1 for correct answer)
3 a Boxer sample 1 = 75%; sample 2 = 45%; sample 3 = 58%
Labrador sample 1 = 5%; sample 2 = 5%; sample 3 = 3% (4 marks)
b Boxer mean = 59%; Labrador mean = 4% (2 marks)
c <2 months (1 mark)
d Two from the following:
Mitotic index is on the upper limit of the category; (1 mark)
survival >70 months likely; (1 mark)
two counts exceed upper limit for category; (1 mark)
suggests that tumour may become more severe/is entering faster
growth stage. (1 mark)
Definitions

Benign: grows slowly, does not spread far.

Malignant: grows quickly and spreads to other tissues affecting function

Metastasis: tumour spreads to blood and lymph, and moves through the body causing secondary tumours.
Bacteria always reproduce asexually, using binary fission ("splitting in two").

1 The DNA in a bacterium is always circular (or rather a closed loop), and is often attached to an invagination of
the cell membrane.

2 Before cell division the DNA is replicated. Unlike eukaryotic cells, there are no histone proteins, and the DNA
is not coiled into chromosomes.

3 The cell elongates from the middle, separating the two DNA molecules, which are attached to different parts of
the membrane.

4 A new cell wall, or septum, is formed down the middle of the elongated cell.
5 Eventually the septum meets, dividing the cell in two.
Although this looks a bit like mitosis in eukaryotic cells, it isn't. There is no nucleus, no chromatids, no centrioles
and no spindle. Binary fission is very fast and bacteria can double every 10 min under optimum conditions,
though the doubling time is usually slower.

Bacterial Spores
Many bacteria can produce spores. These are specialised dormant cells formed in response to adverse conditions,
and they can survive long periods of high temperatures (even boiling), desiccation, low nutrients, radiation and
chemicals. When conditions return to normal the spores "germinate" and develop into normal cells. Although
spores are a little like plant seed they are not used for reproduction. Because they are so difficult to kill, spores
can cause problems for sterilisation.
Viruses can only reproduce inside host cells. The general strategy is to use the host cell enzymes to replicate and translate viral DNA, making more virus
particles, which then burst out of the cell.

The mRNA is now used to synthesise more viral

The virus attaches to the host cell RNA and capsid proteins.
membrane, which stimulates
endocytosis.

The RNA and proteins are assembled into new

The viral envelope fuses with the vesicle virus particles (without envelopes). The glycoproteins
membrane, releasing the nucleocapsid migrate to the cell membrane.
into the cytoplasm.

The virus particles are released by exocytosis,

The RNA enters the nucleus, where it is collecting the lipid envelope from the cell membrane.
replicated to form mRNA, using the viral This kills the host cell
RNA polymerase enzyme.
Specification

The basic structure of all cell membranes, including cell-surface membranes and the membranes
around the cell organelles of eukaryotes, is the same.

The arrangement and any movement of phospholipids, proteins, glycoproteins and glycolipids in the
fluid-mosaic model of membrane structure. Cholesterol may also be present in cell membranes
where it restricts the movement of other molecules making up the membrane.

Movement across membranes occurs by:

simple diffusion (involving limitations imposed by the nature of the phospholipid bilayer)
facilitated diffusion (involving the roles of carrier proteins and channel proteins)
osmosis (explained in terms of water potential)
active transport (involving the role of carrier proteins and the importance of the hydrolysis
of ATP)
Co-transport (illustrated by the absorption of sodium ions and glucose by cells lining the
mammalian ileum).

Cells may be adapted for rapid transport across their internal or external membranes by an increase
in surface area of, or by an increase in the number of protein channels and carrier molecules in, their
membranes.
Students should be able to:

explain the adaptations of specialised cells in relation to the rate of transport across their
internal and external membranes
explain how surface area, number of channel or carrier proteins and differences in gradients
of concentration or water potential affect the rate of movement across cell membranes.

Previous knowledge
Role of the cell membrane
Definition of diffusion and factors that affect the rate of diffusion
Hydrophilic and hydrophobic
Phospholipid structure

Practicals
Osmosis investigation
Effect of alcohol on cell membrane permeability

Biofact sheets: 8, 116, 128, 225,

 Phospholipid structure
Phospholipid consists of glycerol;
To which are joined two fatty acids;
And a phosphate;
formed by condensation reaction
Phosphate head is hydrophilic/polar
Fatty acid tail is hydrophobic

Closer look at the plasma proteins

Intrinsic proteins molecules pass through entire bilayer

Membrane structure
Described as fluid mosaic:
Fluid: molecules within the membrane able to move;
Mosaic: mixture of phospholipid and protein
Double layer of phospholipid molecules (described across the page)
In the membrane the phospholipids are arranged as bilayer in membrane;
hydrophilic heads on the outside and hydrophobic tails on the inside;
The nature of the fatty acid chains can change the properties of the
membrane like fluidity. Unsaturated fatty acids have a lower melting
point, so are more prominent in membranes in colder environments
Some of the proteins have channels/pores;
Some have specific binding sites and are carrier proteins
To move molecules passively by facilitated diffusion or through the hydrolysis of
ATP in active transport.
Extrinsic proteins only in one layer
Those on the outer side often act as receptors for hormones or other chemicals,
can act as enzymes, antigens. Those on the inner side can also act as enzymes or
attach to the cytoskeleton
Many of the proteins and phospholipids have carbohydrates attached forming
glycolipids and glycoproteins that make up the Glycocalyx
The membrane also contains
cholesterol which helps to maintain fluidity, preventing interactions between
fatty acid chains
Internal membranes and compartmentalisation

All reactions occurring in cells take place in certain space compartment, which is separated from other compartments by means of semipermeable membranes. They help
to separate even chemically quite heterogeneous environments and so to optimize the course of chemical reactions.

Lysosomes are membrane bound organelles, sealing hydrolytic enzymes and this prevents an unwanted auto-digestion of other organelles within cell.

It allows the organization of enzymes in sequence so that reactions can occur more rapidly as products of one reaction form the substrate for the next reaction and are
already close to the enzyme. It makes it more likely for enzyme substrate complexes to form.

It allows different metabolic processes to occur simultaneously making the cell more efficient

Membranes can be folded to increase the surface area for reactions like the cristae in mitochondria

Pathway for Intracellular transport (ER shows this)

Compartmentalise regions, provides local environments that allows metabolic processes to occur simultaneously that would be incompatible. These membranes will
contain certain enzymes that catalyse the reactions in catabolic and anabolic processes or isolate potentially harmful substances to certain areas.
o SER contains enzymes for steroid and lipid production,
o RER contains enzymes for protein synthesis
o Golgi packaging and distribution of extracellular compounds (e.g hormones)
o Vesicles on lysosomes keep hydrolytic enzymes contained providing a safe area for the destruction of waste or foreign material

Despite its advantages, compartmentalization at the same time puts greater demand on the energy consumption. It arises from a frequent need to use ATP-dependent
transporters, transporting substances across membranes against the concentration gradient and thus creating different environments in different compartments.

Explain how three features of a plasma membrane adapt it for its functions.
1. Phospholipid bilayer:. Forms a barrier to water soluble / charged substances / allows non-polar substances to pass
2. compartmentalisation: increases efficiency of cell as different processes can occur simultaneously in optimum conditions
3. Bilayer is fluid: Can bend to take up different shapes for phagocytosis / form vesicles / self-repair;
4. Channel proteins (through the bilayer)/intrinsic protein: Let water soluble/charged substances through / facilitated diffusion;
5. Carrier proteins (through the bilayer): Allow facilitated diffusion and active transport of specific polar molecules
6. Surface proteins / extrinsic proteins, glycoproteins / glycolipids: involved in cell recognition / act as antigens / receptors/enzymes
7. cholesterol: regulate fluidity
How materials move across membranes

Cell membranes are a barrier to most substances, and this property allows materials to be concentrated inside cells, excluded from cells, or simply separated from the
outside environment.

This is compartmentalisation is essential for life, as it enables reactions to take place that would otherwise be impossible. Eukaryotic cells can also compartmentalise
materials inside organelles.

Obviously materials need to be able to enter and leave cells, and there are five main methods by which substances can move across a cell membrane:

1. Diffusion 2. Passive Transport (facilitated diffusion) 3. Osmosis 4. Active Transport 5. Vesicles

Transport of substances across a membrane by a (specific) trans-membrane protein molecule. So

Net movement of particles from an area of high concentration to
an area of low concentration
Rate determined by
SA, temperature, size of particle, nature of particles (polar/non
polar)
A few substances can diffuse directly through the lipid
bilayer part of the membrane if they are lipid-soluble
molecules such as steroids, vitamins A, D, E and K, or very
small molecules, such as H2O, O2 and CO2.
substances can only cross a membrane if it contains the appropriate protein. There are two kinds of
transport protein:
Channel Proteins form a water-filled pore or channel in the membrane. This allows charged
substances (usually ions) to diffuse across membranes. Most channels can be gated (opened or
closed), allowing the cell to control the entry and exit of ions.
Carrier Proteins have a binding site for a specific solute and constantly flip between two states so
that the site is alternately open to opposite sides of the membrane. The substance will bind on the
side where it at a high concentration and be released where it is at a low concentration.
Active transport is the pumping of substances across a membrane by a trans-membrane protein pump molecule. The protein binds a molecule of the
substance to be transported on one side of the membrane, changes shape, and releases it on the other side. The proteins are highly specific, so there
is a different protein pump for each molecule to be transported. The protein pumps are also ATPase enzymes, since they catalyse the splitting of ATP ..
ADP + phosphate (Pi), and use the energy released to change shape and pump the molecule. Pumping is therefore an active process, and is the only
transport mechanism that can transport substances up their concentration gradient.

Moves molecules against the concentration gradient (low to high)

Using a specific carrier protein

The molecule binds to the protein

ATP is used to cause a conformational change in the shape of the carrier to move the molecule
 A concentrated solution, a solution rich with solute has a low water
Osmosis
potential.
Net movement of water
A dilute solution, a solution with a low solute concentration has a high water
From a high water potential () to a low water potential
potential.
Through a semi permeable membrane
Water always moves from a high to low water potential.
Water can move through the membrane freely as it is a small molecule.
The water potential of pure water is 0. As we add solute the solution
However, when solutes are added to a solution, water surrounds them, interacting with
becomes more negative. So we say that..
the charged regions, forming hydration shells. Water is then unable to move freely. The
Water moves from the less negative to the more negative water potential.
potential for water to move from that region has been reduced thus
Examples of osmosis in living things
Uptake of water by root (hair)
Movement from cell to cell within plant
Guard cell function
Maintain turgor
Water absorption in the large intestine
Reabsorption of water from the nephron or collecting duct in the kidney
100% pure water has = 0, which is the highest possible water potential, so
all solutions have < 0, and you cannot get > 0.

Cells and Osmosis. The concentration of the solution that surrounds a cell will
affect the state of the cell, due to osmosis. There are three possible
concentrations of solution to consider:

Isotonic solution a solution of equal concentration to a cell

Hypertonic solution a solution of higher concentration than a cell
Hypotonic solution a solution of lower concentration than a cell
Cholera is a bacteria that produces toxins which increase secretion of
chloride ions from intestinal epithelial cells into the lumen of the intestine.
Explain why this results in severe diarrhoea (watery faeces) (3)
Water moves into gut from intestinal cells;
By osmosis
Because there is a lower/more negative water potential in the intestinal
lumen
The water potential of a plant cell is 400 kPa. The cell is put in a solution with
a water potential of 650 kPa. Describe and explain what will happen to the
cell. (use diagrams on previous page to help you) (3)
Membrane pulls away from cell wall (cell plasmolysed);
Water leaves the cell by osmosis
Water moves from less negative to the more negative water potential
Some people are lactose intolerant because they do not produce enough lactase
enzyme in the small intestine. Lactose accumulates in the intestines and either
remains unhydrolysed or is converted to other soluble substances by bacteria in the
intestine. Explain how this could lead to diarrhoea in a lactose-intolerant individual.
(2)
Water potential made lower / more negative;
Water enters gut by osmosis
Describe and explain the effect on the cloroplast when it is placed it in a hypotonic
solution. (4)
Organelle swells/increase in pressure and bursts (lysis) so the membranes break down
Water moves into organelle/chloroplast by osmosis
Water moves from the less negative water potential of surrounding solution to the
more negative solution inside the chloroplast
 The processes described so far only apply to small
molecules. Large molecules (such as proteins,
polysaccharides and nucleotides) and even whole cells
are moved in and out of cells by using membrane
vesicles.

Endocytosis is the transport of materials into a cell. Materials are

enclosed by a fold of the cell membrane, which then pinches shut to
form a closed vesicle. Strictly speaking the material has not yet
crossed the membrane, so it is usually digested and the small product
molecules are absorbed by the methods above. When the materials
and the vesicles are small (such as a protein molecule) the process is
known as pinocytosis (cell drinking), and if the materials are large
(such as a white blood cell ingesting a bacterial cell) the process is
known as phagocytosis (cell eating).

Exocytosis is the transport of materials out of a cell. It is the

exact reverse of endocytosis. Materials to be exported must
first be enclosed in a membrane vesicle, usually from the RER
and Golgi Body. Hormones and digestive enzymes are secreted
by exocytosis from the secretory cells of the intestine and
endocrine glands.
Co-transport (Secondary active transport)

In this case of active transport, the energy released from ATP is

not directly used to transport the particular molecule (like
glucose). Instead, it is used to transport other substances (e.g.
sodium cation), which causes the formation of the concentration
gradient across the membrane. It is this gradient that drives the
transport of the (relevant) substance with the help of other
transporters (e.g. SGLT Sodium Glucose Transporter).

1. The co transport, antiport protein, the sodium-potassium pump, pumps sodium ions out of the epithelial cell and potassium ions into the cell, using the energy from
the hydrolysis of ATP to do so. So there is a large build-up of sodium ions in the lumen of the gut.

2. The facilitated diffusion symport protein is the sodium-glucose co-transporter protein, which is only found in the membrane of the epithelial cells of the ileum. This
carrier protein has two binding sites: one for glucose and one for sodium ions, and both molecules must be carried together. Since this process is diffusion, the molecules
are only carried down their concentration gradients. But the very large concentration gradient of sodium ions across the epithelial cell membrane drives the sodium-
glucose co-transporter in one direction only carrying both molecules into the cell. So while the sodium ions are diffusing down their concentration gradient, the glucose
molecules can be carried up their concentration gradient.

4. The glucose diffuses through the epithelial cell and diffuses into the tissue fluid of the villus by facilitated diffusion, through a glucose carrier protein that is only found
on the inner surface of the epithelial cell.

5. The glucose enters the blood capillary by diffusing through gaps between the capillary endothelial cells. The glucose is carried in the blood to every cell in the body,
where it is used for respiration.
 The proteins usually span from one side of the phospholipid bilayer to
the other (integral proteins), but can also sit on one of the surfaces
(peripheral proteins). They can slide around the membrane very
quickly and collide with each other, but can never flip from one side to
the other. The proteins have hydrophilic amino acids in contact with
the water on the outside of membranes, and hydrophobic amino acids
in contact with the fatty chains inside the membrane.

The phospholipids are arranged in a bilayer (i.e. a double layer), with their
polar, hydrophilic phosphate heads facing out towards water, and their
non-polar, hydrophobic fatty acid tails facing each other in the middle of
the bilayer. This hydrophobic layer acts as a barrier to most molecules,
effectively isolating the two sides of the membrane. Different kinds of
membranes can contain phospholipids with different fatty acids, affecting
the strength and flexibility of the membrane, and animal cell membranes
also contain cholesterol linking the fatty acids together and so stabilising
and strengthening the membrane.

Explain how glucose is absorbed in the small
intestine
Sodium removed (from epithelial cell) by active
transport via the sodium- potassium pump;
Into the intestinal lumen;
maintaining sodium concentration gradient
(between lumen
and epithelial cell);
Glucose moves in into the epithelial cell with sodium
Via a specific symport carrier protein
Glucose moves out of the epithelial cell;
By (facilitated) diffusion;
Describe the structure of the cell membrane Describe how the membrane regulates the
movement of molecules across it
Double layer of phospholipid molecules;
Arranged as bilayer in membrane; Non-polar/lipid soluble molecules move through
Head/phosphate hydrophilic/polar and tail/fatty acid phospholipid layer/bilayer;
hydrophobic/non-polar;
Intrinsic proteins/protein molecules passing right through; Small molecules/water/gases move through
Some with channels/pores; phospholipid layer/bilayer;
Extrinsic proteins/proteins only in one layer/on surface; Ions/water soluble substances move through
Molecules can move in membrane/dynamic/membrane
channels in proteins;
contains
cholesterol; Some proteins are gated;
Glycocalyx/carbohydrates attached to lipids/proteins; Reference to diffusion;
Carriers identified as proteins;
Carriers associated with facilitated diffusion;
The nature of the membrane varies depending on the
location and role of cells. Intestinal epithelia have folded
membrane to increase surface area. They contain only
the sodium glucose transport carrier. Certain cells will
possess receptors for hormones that other cells will not.
So whilst the basic structure is the same the constitution
can vary slight form organism to organism and organ to
organ.
Q7. Read the following passage.

The plasma membrane plays a vital role in microorganisms. It forms a barrier

between the cell and its environment, controlling the entry and exit of solutes. This
makes bacteria vulnerable to a range of antiseptics and antibiotics

When bacteria are treated with antiseptics, the antiseptics bind to the proteins in the
5 membrane and create tiny holes. Bacteria contain potassium ions at a concentration
many times that outside the cell. Because of the small size of these ions and their
concentration in the cell, the first observable sign of antiseptic damage to the
plasma membrane is the leaking of potassium ions from the cell. Some antibiotics
damage the plasma membrane in a similar way. One of these is tyrocidin. This is a
cyclic polypeptide consisting of a ring of ten amino acids
10 Tyrocidin and other polypeptide antibiotics are of little use in medicine.

Other antibiotics also increase the rate of potassium movement from cells. It is
thought that potassium ions are very important in energy release and protein
synthesis, and a loss of potassium ions would lead to cell death. Gramicidin A coils
to form a permanent pore passing through the plasma membrane. This pore enables
potassium ions to be conducted from the
15 inside of the cell into the surrounding medium. Vanilomycin also facilitates the
passage of
potassium ions from the cell. A molecule of vanilomycin forms a complex with a
potassium ion and transports it across the membrane. The potassium ion is released
on the outside and the vanilomycin is free to return and pick up another potassium
ion. Vanilomycin depends on the fluid nature of the plasma membrane in order to
function.

20 Polyene antibiotics have flattened ring-shaped molecules. The two sides of the ring
differ from each other. One side consists of an unsaturated carbon chain. This part
is strongly
hydrophobic and rigid. The opposite side is a flexible, strongly hydrophilic region. It
has been shown that polyene antibiotics bind only to sterols. Sterols are lipids found
in the membranes of eukaryotes but not in the membranes of prokaryotic
organisms. It is thought that several
25 sterol-polyene complexes come together. The plasma membranes of eukaryotic cells
treated with these polyene antibiotics lose the ability to act as selective barriers and
small ions and molecules rapidly leak out

Use information in the passage and your own knowledge to answer the
questions.

(a) (i) By what process do potassium ions normally enter a bacterial cell?
Explain the evidence for your answer.

.............................................................................................................

.............................................................................................................

.............................................................................................................

.............................................................................................................(2)
 (ii) Use Ficks law to explain why leakage of potassium ions occurs following
antiseptic damage to the plasma membrane (lines 7 - 8).

.............................................................................................................

.............................................................................................................

.............................................................................................................

.............................................................................................................(2)

(b) (i) Draw a peptide bond showing how the COOH group of one amino acid
joins to the NH2 group of another.

(1)

(ii) How many peptide bonds are there in a molecule of tyrocidin (lines 9 -
10)?

.............................................................................................................(1)

(c) Experiments have shown that vanilomycin is unable to transport potassium ions
across a membrane when it is cooled. Gramicidin A continues to facilitate the
movement of potassium ions at these low temperatures. Explain these results.

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................

...................................................................................................................... (3)

(d) Draw a simple diagram of one of the phospholipid layers to show how polyene
antibiotics allow small ions and molecules to leak rapidly through a plasma
membrane. Use the following symbols to represent the different molecules.

Note that the zigzag line on the symbol for the polyene antibiotic represents its
hydrophobic region.
 (2)
(Total 11 marks)

Answers

M7. (a) (i) Active transport;

Low to high concentration / against concentration gradient;
Reject answers relating only to high concentration in cell
2

(ii) Rate of movement / diffusion proportional to concentration gradient/

difference in concentration;
High concentration of potassium ions inside cell compared to outside;
Must mention high concentration. Ignore reference to other factors if
reasoning is appropriate.
2

(b) (i) O
||
CN
|
H;
1

(ii) 10;
1

(c) Action of vanilomycin depends on fluidity of membrane;

Fluidity reduced / not fluid at low temperatures;
Pore formed by gramicidin A remains in place / permanent;
3

(d) Pore between sterol molecules lined with polyene antibiotic;

Hydrophobic region next to sterol;
2
[11]
Q10. Read the following passage (movement across membranes)

Cholera is a water-borne disease. It is caused by a bacterium. The bacterium

produces a toxin which acts on the epithelial cells of the small intestine and
causes changes in membrane permeability. The cholera toxin affects the
movement of ions through the intestinal wall. It causes the loss of chloride ions
from the blood into the
5 lumen of the small intestine. This prevents the movement of sodium ions from the
lumen of the small intestine into the blood. The resulting high concentration of
ions causes diarrhoea.

Vaccination can produce immunity to cholera. A new vaccine appears to provide

better immunity and has fewer side effects than previously available vaccines.
This
10 vaccine is taken orally. For long-term immunity, a booster dose is required after
two years.

Use information from the passage and your own knowledge to answer the following
questions.

(a) The cholera toxin only affects the epithelial cells of the small intestine (line 2).
Suggest why.

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................3)

(b) (i) Sodium ions normally enter the blood from cells of the intestinal wall
against a concentration gradient. Describe how.

.............................................................................................................

.............................................................................................................

.............................................................................................................

.............................................................................................................

.............................................................................................................(3)

(ii) The high concentration of ions in the small intestine of a person with
cholera causes diarrhoea. (lines 6-7). Explain why.

.............................................................................................................

.............................................................................................................

.............................................................................................................(2)
 (c) The new vaccine for cholera is taken orally (line 10) but some vaccines are not
taken orally. Suggest one reason why some vaccines are not taken orally.

......................................................................................................................

......................................................................................................................(1)

(d) A booster dose of vaccine is required to provide long-term immunity. Suggest

why.

......................................................................................................................

...................................................................................................................... (1)
(Total 10 marks)

M10. (a) Receptor;

Reference to tertiary structure of protein;
Complementary shape;
Q Do not credit same shape but allow suitable
description of complimentary shape.
3

(b) (i) Active transport;

Using ATP;
Carrier proteins;
3

(ii) Water potential lowered in small intestine;

Osmotic loss of water;
QWC First mark only credited if water potential is
clearly
linked to intestine or blood
2

(c) Broken down by enzymes / not absorbed as molecules are too large;
1

(d) Ensures memory cells produced;

1
[10]
Q12. Read the following passage. (movement across membranes and biochemistry)

Human milk contains all the nutrients a young baby needs in exactly the right
proportions. It is formed in the mammary glands by small groups of milk-
producing cells.
These cells absorb substances from the blood and use them to synthesise the
lipids, carbohydrates and proteins found in milk. Milk-producing cells are
roughly cube-shaped
5 and have a height to breadth ratio of approximately 1.2 : 1.

The main carbohydrate in milk is lactose. Lactose is a disaccharide formed by

the condensation of two monosaccharides, glucose and galactose. (A molecule
of galactose has the same formula as a molecule of glucose the atoms are
just arranged in a different way.)

10 Lactose is synthesised in the Golgi apparatus and transported in vesicles

through the cytoplasm. Because lactose is unable to escape from these
vesicles, they increase in diameter as they move towards the plasma
membrane. The vesicle membranes fuse with the plasma membrane and the
vesicles empty their contents out of the cell.

Use the information from the passage and your own knowledge to answer the
following questions.

(a) (i) The breadth of a milk-producing cell is 26 m. Calculate the height of this
cell.

Height = .......................... m (1)

(ii) Describe and explain how you would expect the height to breadth ratio of
an epithelial cell from a lung alveolus to differ from the height to breadth
ratio of a milk-producing cell.

.............................................................................................................

.............................................................................................................

.............................................................................................................

.............................................................................................................(2)

(b) How many oxygen atoms are there in a molecule of

(i) galactose;

.............................................................................................................(1)

(ii) lactose?

.............................................................................................................(1)
(c) The lactose-containing vesicles increase in diameter as they move towards the
plasma membrane of the milk-producing cell (lines 11-12). Use your knowledge
of water potential to explain why.

......................................................................................................................

......................................................................................................................

......................................................................................................................

...................................................................................................................... (2)

(d) Suggest one advantage of milk-producing cells containing large numbers of

mitochondria.

......................................................................................................................

......................................................................................................................

......................................................................................................................

...................................................................................................................... (2)

(e) Some substances pass through the plasma membrane of a milk-producing cell
by diffusion. Describe the structure of a plasma membrane and explain how
different substances are able to pass through the membrane by diffusion.

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................(6)
(Total 15 marks)
M12. (a) (i) 31/31.2;
1

(ii) Ratio would be less/smaller;

Cell is thin / has large surface area / (adapted) for diffusion;
Accept converse. Must relate to concept of ratio.
2

(b) (i) 6;
1

(ii) 11;
1

(c) Water potential inside vesicle more negative/lower;

Water moves into vesicle by osmosis/diffusion;
2

(d) Mitochondria supply energy/ATP;

For active transport / absorption against concentration
gradient / synthesis / anabolism / exocytosis / pinocytosis;
Do not credit references to making,
creating or producing energy.
2

(e) 1 Phospholipids forming bilayer/two layers;

2 Details of arrangement with heads on the outside;
3 Two types of protein specified;
e.g. passing right through or confined to one layer /
extrinsic or intrinsic /
channel proteins and carrier proteins /
two functional types
4 Reference to other molecule e.g. cholesterol or glycoprotein;
5 Substances move down concentration gradient/from high to low
concentration;
Reject references to across or along a gradient
6 Water/ions through channel proteins/pores;
7 Small/lipid soluble molecules/examples pass between phospholipids/
through phospholipid layer;
8 Carrier proteins involved with facilitated diffusion;
Ignore references to active transport.
Credit information in diagrams.
max 6
[15]
Questions
1 Draw a labelled diagram to show the arrangement of molecules in the
fluid-mosaic model of the membrane structure. Indicate the outside
and the inside of the cell on your diagram.

2 Explain why this model of the membrane structure is known as the fluid-mosaic model.

3 What are the functions of the components of cell-surface membranes?

4 Diffusion is the net movement of molecules or ions from an area of higher concentration to
an area of lower concentration.
a Explain what is meant by net movement.
b Explain how each of the following factors affects the rate of diffusion.
i The concentration gradient.
ii The area of the exchange surface.
iii The thickness of the exchange surface.
iv The size and nature of the diffusing molecules.

5 Facilitated diffusion is a special type of diffusion enabling molecules, such as glucose; and
amino acids, and charged particles, such as sodium ions, to diffuse into and out of cells.
a Explain how facilitated diffusion differs from simple diffusion.
b In what ways are the two processes similar?
c By means of labelled diagrams, show how large molecules enter cells by facilitated
diffusion.

6 Osmosis is the diffusion of water molecules through partially permeable membranes. It is

defined in terms of water potential.
a Complete the following passage by filling in the missing words or phrases.
Osmosis is the diffusion of water from an area of .................... or .................... water
potential to an area of .................... or .................... water potential through a partially
permeable membrane. Pure water has a water potential of ..................... Adding a solute
to pure water .................... the water potential. The more solute that is added, the more
.................... the solution becomes and the value of the water potential is more
..................... Two solutions that have the same water potential are said to be
.....................
b Explain what happens when (i) red blood cells and (ii) discs of potato tuber tissue are
placed in pure water.

7 Explain how active transport differs from passive forms of transport.

8 Describe how energy from ATP is used in active transport.

Answers to questions
9 Accept diagrams which show a phospholipid bilayer with the molecules represented accurately with their
heads and tails oriented correctly. There should be proteins in the membrane look for extrinsic and
intrinsic proteins, cholesterol molecules, glycolipid and glycoproteins attached to the extrinsic proteins.
Credit some attempt to represent channel proteins and carrier proteins. The outside and the inside of the
cell should be labelled.

10 The fluid property is due to the fact that the phospholipid molecules move relative to each other, providing
the membrane with flexibility. The protein molecules form the mosaic. The protein molecules vary in size
and shape.

11 Phospholipid bilayer allows lipid-soluble molecules through but prevents entry of water-soluble ones. It
also makes the membrane flexible.
Membrane (intrinsic) proteins span the membrane and allow the passage of larger molecules or ions that
are water-soluble. Some act as carrier proteins to transport the water-soluble substances across the
membrane. Others are channel proteins, facilitating the active transport of sodium and potassium ions, for
example, across the membrane.
Glycolipids and glycoproteins attached to some extrinsic proteins act as receptors and enable cell
recognition.
Proteins provide structural support.
Cholesterol molecules reduce the fluidity and give strength.

12 Diffusion is the net movement of molecules or ions from an area of higher concentration to an area of
lower concentration.
a Molecules in a gas or liquid are moving randomly all the time and will diffuse both ways. Where there is
a difference in concentration, however, there will be a tendency for more molecules to move away from
the higher concentration to where they are in a lower concentration, i.e. down a concentration gradient,
until equilibrium is reached and there is an even distribution.
Note: Reference should be to down the concentration gradient, not along or across.
b Explain how each of the following factors affects the rate of diffusion:
The concentration gradient: the greater the concentration gradient, the faster the rate of diffusion. As
the difference in concentration becomes less, the rate will slow down.
The area of the exchange surface: the larger the surface area, the faster the rate of diffusion. In the
human body, microvilli increase the surface area of the cell-surface membranes of cells involved in
absorption.
The thickness of the exchange surface: the thicker the exchange surface, the slower the rate of
diffusion as the particles have a longer distance to travel. Thin exchange surface membranes are the
most efficient.
The size and nature of the diffusing molecules: small molecules diffuse faster than large ones. Fat-
soluble molecules diffuse faster than water-soluble ones. Polar molecules diffuse faster than non-polar
molecules. Oxygen, however, is non-polar and diffuses quickly. As the degree of polarity increases, the
rate of diffusion decreases. Water is polar and small enough, however, to get through quickly. So size
and polarity are not mutually exclusive.

13 Facilitated diffusion is a special type of diffusion enabling molecules, such as glucose and amino acids,
and charged particles, such as sodium ions, to diffuse into and out of cells.
a Simple diffusion is the movement of substances through a cell-surface membrane. Facilitated diffusion
enables larger molecules and charged particles to diffuse through cell-surface membranes. It involves
the carrier proteins and protein channels.
b Both processes are passive, i.e. do not use energy. In both, movement is down a concentration
gradient. The rates of both depend on the surface area of the exchange surface, the thickness of the
exchange surface and the nature of the diffusing molecules.
c Accept suitable diagrams showing proteins which span the cell-surface membrane, higher
concentration of molecules outside the cell and lower concentration inside. Credit diagrams of channel
proteins and carrier proteins.

14 Osmosis is the diffusion of water molecules through partially permeable membranes. It is defined in terms
of water potential.
 a Osmosis is the diffusion of water from an area of higher or less negative water potential to an area of
lower or more negative water potential through a partially permeable membrane. Pure water has a
water potential of zero. Adding a solute to pure water lowers the water potential. The more solute that
is added, the more concentrated the solution becomes and the value of the water potential is more
negative. Two solutions that have the same water potential are said to be isotonic.
b i Red blood cells burst because they have a lower or more negative water potential and water is taken
up by osmosis. The cells swell and burst because the cell-surface membrane cannot stretch to
accommodate the water.
ii Discs of potato tuber become turgid but the cells do not burst. Water enters the cells by osmosis
because the cells have a lower or more negative water potential. The cell contents swell and push up
against the cellulose cell walls which then exert an inward pressure preventing the cells from bursting
and from taking up any more water.

15 In active transport, molecules are moved from a lower concentration to a higher concentration against a
concentration gradient. This process requires energy. It involves the use of carrier proteins in a similar way
to facilitated diffusion, in that the molecules attach to the carrier protein, the protein changes shape and the
molecule moves across the membrane into the cell. Energy from ATP is needed to move the molecule
against the concentration gradient.
16 ATP attaches to the carrier protein on the inside of the cell causing it to change shape or position in the
membrane and to allow molecules passage into the cell. ADP and P are released from the carrier protein
which then reverts to its original receptive shape or position.
Accept from suitably annotated diagrams.
Specification Biofactsheets: 99,40, 100, 169, 112, 162

Each type of cell has specific molecules on its surface that identify it. These molecules include
proteins and enable the immune system to identify:
pathogens
cells from other organisms of the same species
abnormal body cells
toxins.

Definition of antigen. The effect of antigen variability on disease and disease prevention.
Phagocytosis of pathogens. The subsequent destruction of ingested pathogens by lysozymes.
The response of T lymphocytes to a foreign antigen (the cellular response).

The role of antigen-presenting cells in the cellular response.

The role of helper T cells (TH cells) in stimulating cytotoxic T cells (TC cells), B cells and
phagocytes. The role of other T cells is not required.

The response of B lymphocytes to a foreign antigen, clonal selection and the release of monoclonal
antibodies (the humoral response).
Definition of antibody.
Antibody structure.
The formation of an antigen-antibody complex, leading to the destruction of the antigen,
limited to agglutination and phagocytosis of bacterial cells.
The roles of plasma cells and of memory cells in producing primary and secondary immune
responses.

The use of vaccines to provide protection for individuals and populations against disease. The
concept of herd immunity. The differences between active and passive immunity.

Structure of the human immunodeficiency virus (HIV) and its replication in helper T cells.

How HIV causes the symptoms of AIDS. Why antibiotics are ineffective against viruses.

The use of monoclonal antibodies in:

targeting medication to specific cell types by attaching a therapeutic drug to an antibody
medical diagnosis.
Details of the production of monoclonal antibodies is not required.

Ethical issues associated with the use of vaccines and monoclonal antibodies.
The use of antibodies in the ELISA test.
Students should be able to:
discuss ethical issues associated with the use of vaccines and monoclonal antibodies
evaluate methodology, evidence and data relating to the use of vaccines and monoclonal
antibodies.

Previous knowledge
Explain how white blood cells respond to disease
Explain how a vaccine works and why they make us immune
Explain how the body prevents the entry of microbes
Explain what transmission is and give some examples of how disease is transmitted
Explain what a pathogen is and why they make us feel ill
Knowledge of protein structure (primary, secondary, tertiary and quaternary structure)
Know what lysosomes are
 All living cells have antigens as part of their cell membrane or cell wall. Their purpose is for
cell communication, and cells from different individuals have different antigens, while all the
cells of the same individual have the same antigens. Antigens are genetically controlled, so
close relative have more similar antigens than unrelated individuals. Blood groups are an
example of antigens on red blood cells, but all cells have them.

What is an antigen?
Molecule/part of molecule/protein/glycoprotein;
Stimulates immune response;

If antigens and antibodies have a specific relationship and if our bodies have immunological memory How come we
can get the flu every year??

Antigenic variability: microbes have constantly-changing antigens. It is caused by mistakes in DNA or RNA replication
(mutations) due to poor polymerase enzymes.

The immune system

The cells of the immune system are the white blood cells (or leukocytes). Leukocytes are derived
from stem cells, which are produced in huge numbers in the bone marrow (the soft centre of large
bones). These stem cells differentiate to form dozens of different kinds of leukocytes, which fall into
four categories

Pathogens
Disease causing organisms

Cause illness by: Damaging tissues producing toxins

Humans have three lines of defence against invading pathogens:

1. Barriers the skin and associated chemicals stop microbes entering the body
2. The non-specific immune system phagocytes quickly destroy microbes that pass the first line of defence
3. The specific immune system lymphocytes kill any microbes that pass the second line of defence, and remain on
guard for future attacks.
Explain how the defence mechanisms of the body reduce the
chance of entry by a pathogen.

Epidermis of skin is a dead, keratinised (tough protein) layer so

pathogens cannot penetrate;

Mucus in respiratory system is trapping sticky pathogens;

cilia move fluid / mucus removing pathogens;

Tears / saliva / mucus contain lysozyme which breaks down

(lyses) bacterial peptidoglycan cell walls;

Stomach contains hydrochloric acid which destroys bacteria;

blood clot prevents entry;

Respiratory tract contains mucus (containing lysozyme) and

ciliated cells

Fluid nature of tears wash away bacteria;

Vaginal acid destroys bacteria;

Commensal bacteria on skin out-compete pathogen;

Sebum (fatty acid) inhibits bacterial growth;

Commensalism: one organism benefits the other is unaffected

Mutualism: both organisms benefit
Parasitism: one organism benefits the other is harmed
The non-specific immune system kills pathogens quickly and indiscriminately.
Although effective, the non-specific immune system does not "learn from
experience", so it does not lead to immunity to a disease.
The specific immune system is a more complex system that not only kills
invading pathogens, but also remembers the pathogen's features so that it can
be killed quickly on subsequent infections. While all animals have a non-specific
immune system, only vertebrates have a specific immune system, so it must be a
later evolutionary advance. The key difference of the specific immune system is
that it is capable of recognising foreign cells as distinct from its own cells, an
ability called self/nonself recognition. It does this by making use of antigens.
The parts of the immune system are spread all over the body. They include:
The lymph and blood vessels. These transport pathogens and leukocytes all over the
body.
The lymph nodes. These contain millions of phagocyte and lymphocyte cells, which
identify and remove pathogens from lymph.
The spleen. This also contains millions of phagocyte and lymphocyte cells, which
identify and remove pathogens from blood.
The thymus. This is where blood stem cells are differentiated into T-lymphocytes.
 Non-specific chemical response

Complement system: 20 different proteins which kill microbes by making

pores in their membranes and interferons are proteins that can also inhibit
Phagocytosis: Non-specific cellular response
viral reproduction inside cells.
Phagocytes engulf pathogens/microorganisms;
Inflammation: Caused by the release of chemicals like histamine,
Enclosed in a vacuole / vesicle/ phagosome;
prostagladins and kinins which cause vasodilation to increase blood flow to
Fuses with lysomsome to for a phagolysosome
an area (redness), allows capillary leakage so phagocytes can access the area
Lysosomes have enzymes;
(swelling)
That digest/hydrolyse molecules/proteins/lipids/microorganism;
Pyrogen chemicals cause body temperature to increase (fever) 370C 390C
Monocytes mature into macrophages when they move from the blood into
helping to inhibit some pathogenic growth
tissues. They can secrete substances to attract other white blood cells, and
they can also become antigen presenting cells (APCs) and help stimulate the
cell mediated immune response.

Neutrophils: circulate in the blood

 The specific immune system: B There are two kinds of lymphocyte B-lymphocytes (or just B-cells) and T-lymphocytes (or just Tcells). B-cells are called
cells, humoral response that because they mature from stem cells in the bone marrow. T-cells are called that because they mature from stem cells in
the thymus.

B cells are involved in the humoral response. They are Lymphokines/cytokines stimulate the B cells to
produced and mature in the bond marrow divide by mitosis

These will differentiate into

TH-cells bind to the B cell and are
B cell binds to the antigen with the complementary Plasma cells: that secret antibodies that will
activated to release
shape to the antibodies on the membrane. bind to antigens and agglutinate pathogens,
cytokines/lymphokines, these will
neutralise/immobilise pathogens or kill them
stimulate the lymphocytes
It then processes and displays the antigen, becoming an directly by causing cell lysis
antigen presenting cell
Memory cells: these provide immunity, that on
reinfection they quickly divide producing large
quantities of antibodies
A single B cell can divide to form 106 plasma cells; each of these can release 103 antibodies every second
for 4 days!!!!!

T-cells have receptor molecules on their surfaces which are
very similar, but not identical, to antibodies. These
receptors also bind specifically to antigens to form antigen-
receptor complexes. Each T-cell has around 105 receptor
proteins, and again there are about 108 different types of T-
cell, each with slightly different receptor molecules, so they
can also specifically bind to any conceivable antigen. T-cells
do not secrete soluble proteins.
Antibodies
Antibodies are proteins (immunoglobulins), with a specific, complimentary shape to an antigen.
They bind with antigens forming an antigen antibody complex
Protein molecules are particularly well suited to carry out the role of antibodies because they can exist
in a large variety of different shapes; due to various primary structures that will result in a wide variety
of tertiary shapes. They are complementary to specific antigens
Antibodies all have a similar structure composed of 4 polypeptide chains (2 heavy chains and 2 light
chains) joined together by strong disulphide bonds to form a Y-shaped structure.
The stem of the Y is called the constant region because in all immunoglobulins it has the same amino
acid sequence, and therefore same structure.
The ends of the arms of the Y are called the variable regions of the molecule because different
immunoglobulin molecules have different amino acid structure and therefore different structures.
These variable regions are where the antigens bind to form a highly specific antigen-antibody complex
Each B-cell has around 105 membrane-bound antibody molecules on its surface and can also secrete
soluble antibodies into its surroundings. Every human has around 108 different types of B cell, each
making antibodies with slightly different variable regions. . Each antigen contains a variety of binding
points called epitopes. Antibodies can therefore bind specifically to 108 different antigens, so there
will be an antibody to match almost every conceivable antigen that might enter the body
T cells and cell mediated immunity

Lymphokines will cause T cells to start to divide and they will

differentiate into various types of cell

Cytotoxic T cells (TC): bind to antigens on infecting cells and kill them
by releasing perforin proteins that create pores in the cell that lets
water diffuse in and the cells burst

Helper T cells (TH): are stimulated to release chemicals

(lymphokines/cytokines) when they encounter foreign antigens. This
causes other white blood cells to become active, for example mitosis
in B and T cells.

Memory cells: can divide rapidly with exposure to a familiar antigen.

1st exposure only a few lymphocytes for the antigen so it takes time for clonal selection to occur
(clonal selection: a particular lymphocyte is stimulated to divide by mitosis). In this period the
symptoms of the infection are shown

After this response both T and B memory cells remain in the blood. Subsequent infection by the
same antigen stimulates a secondary immune response where there is a paid production of
antibodies, in large quantities. This is immunity.

But this secondary immune response does not always work, due to antigenic variability, where
the antigen has changed slightly.
 Vaccines:

Contain dead or weakened (attenuated) pathogens or antigens

They initiate an immune response and on exposure to an active pathogen the secondary immune
response is stimulated and thus the person does not experience the symptoms of the disease.

Making vaccines

Live non virulent strains: microbes are selected for most non virulent and slowest growing strains

Killed organisms: killed using chemicals or radiation so as not to denature the antigen proteins
Types of immunity
Isolated antigens
Passive immunity: injections of antibodies
Modified toxins
Active immunity: when an immune response has been initiated
and the persons white blood cells stimulated

Type Active Immunity Passive immunity

Exposure to the pathogen, the body goes through Achieved when antibodies are passed across the
Natural the primary immune response, and the person placenta or in the colostrum (the milk produced in
gets ill the first few days after birth

Injection of attenuated pathogens, modified

Artificial Injections of antibodies
toxins or antigens
 Herd immunity (also called herd effect, community immunity,
population immunity, or social immunity) is a form of indirect
Active Immunity Passive Immunity protection from infectious disease that occurs when a large
percentage of a population has become immune to an infection,
No exposure to antigen thereby providing a measure of protection for individuals who
Exposure to antigen
required are not immune. It disrupts the transmission of the disease,
often this can be a very high number like 95% of the population
Slow to develop protection Immediate protection before it is effective, and with illegal immigration, refugees, poor
response to vaccines from individuals, people living it cramped
Memory cells produced No memory cells conditions, antigenic variability difficult to provide to all
vulnerable people at the right time, expensive to make, animal
testing, unkonwn side effects (chrons disease, autism linked to
Long term protection Short term protection
MMR etc

Ethics of vaccines
HIV (human immunodeficiency virus)

AIDS (Acquired Immune deficiency syndrome)

The human immunodeficiency virus (HIV) is a lipid membrane enveloped

retrovirus. The membrane not made by the virus itself, but derived from a
host cell membrane.

It comprises 2 copies of single-stranded RNA together with some enzymes,

surrounded by an icosahedral capsid, which is in turn surrounded by a
sphere of matrix proteins attached to a lipid envelope.

A retrovirus is one that contains single stranded RNA and the enzyme
reverse transcriptase
Transmission Through infected semen or vaginal secretions during sexual activity,
or through infected blood in transfusions or contaminated needles. Infected
mothers can also pass the virus on to their children through the placenta or milk.
Before 1985 many hospital patients, especially haemophiliacs, became infected
through blood transfusions, but since 1985 all blood donations in the UK are
tested for HIV. Many drug addicts have been infected through sharing needles.
By far the most important method of transmission of HIV world-wide is
unprotected sexual intercourse. HIV cannot survive in air and therefore cannot
be transmitted by skin contact or kissing.
Signs and Symptoms
Like other retroviruses, HIV has a long latency of 8-10 years, during which time
there are no symptoms, but the individual is infectious. After this period the
person starts to shows mild symptoms of the AID-related complex (ARC), such as
tiredness, fever, weight loss and diarrhoea. This is followed by the more serious
symptoms of full-blown AIDS. Since the immune system no longer functions the
patient has no defence against a variety of opportunistic infections. The most
common of these are Kaposi's sarcoma (a skin cancer), TB and pneumonia, which
is usually fatal.
HIV does not become AIDS immediately; time between HIV positive and AIDs
varies. (depending on things like age and access to healthcare and the strain of
the virus). A person has AIDs when symptoms of the immune system failure
begin. Initially affecting the mucus membranes, progression to TB, Chronic
diarrhoea and bacterial infections and culminating in more serious
Antibiotics kill bacteria by interfering with metabolic processes. Viruses are
acellular, do not have their own metabolic processes and organelles and make
use of human enzymes and ribosomes, thus antibiotics will not target them
Infection mechanism
HIV in the blood attaches to cells that carry the "CD4" antigen, including the T
lymphocyte and macrophage white blood cells. After entering these cells it becomes a
provirus in the nuclear DNA, remaining dormant but being replicated for a long latency
period of 8-10 years. Eventually the virus particles are re-assembled and emerge into the
blood, rupturing and killing the T cells in the process. The lack of T cells leaves the
immune system severely compromised.
It targets TH cells as they are the cells with the complimentary receptor and the
destruction of these cells means that the B and T cells cannot be activated to mount an
immune response
Treatment There is as yet no cure or vaccination for AIDS, though drugs like AZT can
delay its onset for many years.
Prevention Vaccinations are difficult because the HIV genome is highly variable (probably
because reverse transcriptase makes many base copying errors). Prevention of AIDS has
concentrated on "safe sex" education (using condoms and reducing promiscuity), not
sharing needles, and screening blood transfusions.
Infection mechanism:
Attaches to host cell, releases the capsid into the cell which includes RNA and enzymes.
Reverse transcriptase converts RNA to cDNA and then the double stranded DNA is inserted
into the human DNA. Host cells will manufacture viral proteins and new viruses are
assembled and eventually bud from the cell.
Monoclonal antibodies Cancer treatment

Reference to hybridomal cells from tumour/cancer cells giving them the Cancer cells have antigens called tumour markers that are not found on normal body cells. So
ability to divide quickly and continuously and B-lymphocytes (plasma monoclonal antibodies can be made to bind to these tumour markers.
cell) producing a particular antibody complementary one antigen
Anticancer drugs can be attached to these antibodies so when they attach to the cancer cell the drug
will accumulate in the vicinity of the tumour minimising the impact on healthy cells.
The fact they can be produced to target any antigen, means that
they have potential in medical diagnosis and treatments.

Diagnostic uses: pregnancy test

Pregnancy tests detect human chorionic gonadotropin (hCG)

found in a pregnant womans urine

The area where urine is applied contains antibodies

complementary to hCG, bound with a blue dye

If hCG is present in the urine it will bind to the antibody

(antigen-antibody complex)

The urine will move up the test strip carrying antibody with it

At the other end of the strip there are immobilised hCG

antibodies

As the antibodies being carried in the urine reach this area, if

they are carrying an hCG they will start to accumulate and the
area will appear blue.
 ELISA (enzyme-linked immunosorbent assay)

This technique is performed to detect the presence

and/or amount of a target protein/antigen of interest.
Or it can be used to detect if a person contains
antibodies for a certain antigen.

This process can be used to detect for allergies or

infection.

An antibody is used which has an enzyme attached to

it; this will catalyse the conversion of a substrate to a
coloured product. The intensity of the colour can be
an indicator of the quantity of antigen/antibody

The antibodies are often generated using animals;

clearly this raises some ethical issues.

Mechanism of the direct ELISA test

Antibody is bound inside the tray (complementary to the antigen under investigation)
Blood sample is added that may or may not contain the antigen
A detection antibody is added that is (complementary to the antigen in question). This antibody contains an attached enzyme.
The tray is washed to remove any unbound antibodies and samples, particularly the antibody attached to the enzyme

If the antigen was present it will be bound to the immobilised antibody and the detection antibody will also bind to it.
Finally a substrate solution is added and any enzyme that has been bound will react with this producing a colour change and a positive result.
Q1. Read the following passage. (Immune system)

Pathogens affect humans. They also affect farm animals. Once pathogens have
entered the body of an animal they can cause disease. Vets sometimes have
difficulty identifying the disease from which a particular animal is suffering. Until
recently, they have had to take blood samples and send them to a laboratory.
The laboratory carries 5 out tests on the sample.

New tests have been developed. Some of these new tests use monoclonal
antibodies. Tests using monoclonal antibodies are fast, specific and allow vets to
identify a disease while they are still on the farm.

Brucellosis is a disease of cattle. It is caused by bacteria. These bacteria can

infect
10 people who drink milk or eat dairy products from infected cattle. A test using
monoclonal antibodies allows vets to identify cattle that are carriers. The carriers
are
cattle that carry the brucellosis bacteria but do not show any symptoms of the
disease.

Use the information from the passage and your own knowledge to answer the
following questions.

(a) Other than bacteria, name one type of pathogen (line 1).

......................................................................................................................(1)

(b) Give two ways in which a pathogen may cause disease when it has entered the
body (lines 12).

1 ...................................................................................................................

2 ................................................................................................................... (2)

(c) Some new tests use monoclonal antibodies (lines 67).

(i) Explain why these antibodies are referred to as monoclonal.

.............................................................................................................

.............................................................................................................(1)

(ii) Tests using monoclonal antibodies are specific (line 7). Use your
knowledge of protein structure to explain why.

.............................................................................................................

.............................................................................................................

.............................................................................................................

.............................................................................................................(3)
 (d) The tests using monoclonal antibodies allow vets to identify brucellosis while they
are still on a farm. Explain the advantages of this.

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................(3)

Q2. Read the following passage.

Campylobacter jejuni is a bacterium. It is one of the commonest causes of

diarrhea in humans. The illness that it causes does not usually last very long and
many sufferers do not even go to the doctor. The only treatment required is the
use of oral rehydration solutions to replace the water lost by diarrhoea. In 1998,
laboratory tests confirmed 5 60 000 cases of diarrhoea caused by this
bacterium in the UK. The bacterium was more frequently found in males than in
females with a ratio of 1.5 : 1.

In rare cases, the nervous system may be affected. Scientists are now beginning
to understand the cause of this. Sugars in the antigens on the surface of the
bacteria are identical to some of the sugars on the surface of nerve cells.
Antibodies produced 10 against the bacteria may therefore attack the bodys
nerve cells. There can be serious problems if this leads to paralysis of the
diaphragm. Breathing difficulties result and the patient may die.

Use information in the passage and your own knowledge to answer the following
questions.

(a) (i) The number of cases of diarrhoea confirmed as being caused by

Campylobacter jejuni in the UK in 1998 was 60 000 (lines 45). Explain
why the true number of cases is thought to be more than this.

.............................................................................................................

.............................................................................................................(1)

(ii) Calculate the number of cases of diarrhoea confirmed as being caused by

Campylobacter jejuni in men in 1998.

Answer ..........................................(1)
 (b) Explain how an oral rehydration solution (ORS) replaces water lost by diarrhoea
(lines 34).

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................

...................................................................................................................... (3)

(c) Explain why antibodies produced against Campylobacter jejuni also attack nerve
cells (lines 9 10).

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................(3)

(d) Explain how paralysis of the diaphragm leads to breathing difficulties (line 11).

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................(2)
(Total 10 marks)

Q3.Read the following passage.

Microfold cells are found in the epithelium of the small intestine. Unlike other epithelial
cells in the small intestine, microfold cells do not have adaptations for the absorption of
food.

Microfold cells help to protect against pathogens that enter the intestine. They have
receptor proteins on their cell-surface membranes that bind to antigens on the surface
of pathogens. 5 The microfold cells take up the antigens and transport them to cells
of the immune system. Antibodies are then produced which give protection against the
pathogen. Scientists believe that it may be possible to develop vaccines that make use
of microfold cells. These vaccines could be swallowed in tablet
form. 10

Use information from the passage and your knowledge to answer the following.
(a) Microfold cells do not have adaptations for the absorption of food (lines 2-3).
Give two adaptations that other epithelial cells have for the absorption of food.

1.....................................................................................................................

2...................................................................................................................... (2)

(b) (i) Microfold cells have receptor proteins on their cell-surface membranes that
bind to antigens (line 5). What is an antigen?

..............................................................................................................

..............................................................................................................

..............................................................................................................1)

(ii) Microfold cells take up the antigens and transport them to cells of the
immune system (lines 6-7). Antigens are not able to pass through the cell-
surface membranes of other epithelial cells. Suggest two reasons why.

...............................................................................................................

...............................................................................................................

...............................................................................................................

...............................................................................................................

...............................................................................................................(2)

(c) Scientists believe that it may be possible to develop vaccines that make use of
microfold cells (lines 9-10). Explain how this sort of vaccine would lead to a
person developing immunity to a pathogen.

...............................................................................................................

...............................................................................................................

...............................................................................................................

...............................................................................................................

...............................................................................................................

...............................................................................................................

...............................................................................................................

...............................................................................................................

............................................................................................................... (5)
(Total 10 marks)
Q4. Read the following passage.

The life cycle of the malarial parasite consists of a number of stages. Some of these stages
occur in humans and some occur in mosquitoes. At each stage, the parasite has different
antigens on the surface of its cells. Attempts have been made to extract some of these
antigens and use them to make vaccines to combat the disease. A trial has recently been
carried out 5 with one of these vaccines. An injection of the vaccine was given to a group
of people chosen at random at the start of the trial. Another injection was given 30 days later.

Blood samples were taken at regular intervals throughout the trial. After the first injection,
the concentration of antibody in the blood rose slowly then fell quickly. After the second
injection, the concentration rose quickly. It reached a maximum concentration of
10 approximately twice the concentration it reached after the first injection.

Use information from the passage and your own knowledge to answer the following
questions.

(a) What is meant by antigens (line 3)?

......................................................................................................................

......................................................................................................................

......................................................................................................................(2)

(b) (i) Use information from the passage to sketch a graph to show the effects of the
two injections on the concentration of antibody in the blood.

(3)

(ii) Suggest one reason why it was necessary to give two injections of the vaccine
(line 6).

.............................................................................................................

.............................................................................................................(1)

(iii) Although this vaccine is made from antigens from malarial parasites, it does
not cause malaria. Explain why this vaccine does not cause malaria.

.............................................................................................................

.............................................................................................................

.............................................................................................................(2)

(c) The blood from those taking part in the trial was also examined under the
microscope at the beginning of the trial. Explain how this would enable those who
had malaria to be identified.

......................................................................................................................

...................................................................................................................... (1)(Total 9 marks)

Q8. Read the passage below. (immunity)

Most cases of cervical cancer are caused by infection with Human Papilloma
Virus (HPV). This virus can be spread by sexual contact. There are many types
of HPV, each identified by a number. Most of these types are harmless but types
16 and 18 are most likely to cause cervical cancer.

A vaccine made from HPV types 16 and 18 is offered to girls aged 12 to 13.Three
5 injections of the vaccine are given over six months. In clinical trials, the vaccine
has proved very effective in protecting against HPV types 16 and 18. However, it
will be many years before it can be shown that this vaccination programme has
reduced cases of cervical cancer. Until then, smear tests will continue to be
offered to
10 women, even if they have been vaccinated. A smear test allows abnormal cells
in the cervix to be identified so that they can be removed before cervical cancer
develops.

The Department of Health has estimated that 80% of girls aged 12 to 13 need to
be vaccinated to achieve herd immunity to HPV types 16 and 18. Herd immunity
is where enough people have been vaccinated to reduce significantly the spread
of HPV through the population.

Use information from this passage and your own knowledge to answer the
following

(a) HPV vaccine is offered to girls aged 12 to 13 (line 5). Suggest why it is offered to
this age group.

......................................................................................................................

......................................................................................................................

...................................................................................................................... (1)

(b) The vaccine is made from HPV types 16 and 18 (line 5). Explain why this vaccine
may not protect against other types of this virus.

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................(2)

(c) Three injections of the vaccine are given (lines 5 to 6). Use your knowledge of
immunity to suggest why.

......................................................................................................................

......................................................................................................................

......................................................................................................................

...................................................................................................................... (2)
 (d) It will be many years before it can be shown that this vaccination programme has
reduced cases of cervical cancer (lines 7 to 9). Suggest two reasons why.

1 ...................................................................................................................

......................................................................................................................

2 ...................................................................................................................

......................................................................................................................(2)

(e) Smear tests will continue to be offered to women, even if they have been
vaccinated
(lines 9 to 10). Suggest why women who have been vaccinated still need to be
offered smear tests.

......................................................................................................................

......................................................................................................................

...................................................................................................................... (1)

(f) Suggest one reason why vaccinating a large number of people would reduce
significantly the spread of HPV through the population (lines 14 to 16).

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................

......................................................................................................................(2)
(Total 10 marks)

Q11. (HIV)
The human immunodeficiency virus (HIV) leads to the development
of acquired immunodeficiency syndrome (AIDS). Eventually, people
5
with AIDS die because they are unable to produce an immune
response to pathogens.

Scientists are trying to develop an effective vaccine to protect

people against HIV. There are three main problems. HIV rapidly
10
enters host cells. HIV causes the death of T cells that activate B
cells. HIV shows a lot of antigenic variability.

Scientists have experimented with different types of vaccine for HIV.

One type contains HIV in an inactivated form. A second type
contains attenuated HIV which replicates in the body but does not
kill host cells. A third type uses a different, non-pathogenic virus to
15
carry genetic information from HIV into the persons cells. This
makes the persons cells produce HIV proteins. So far, these types
of vaccine have not been considered safe to use in a mass
vaccination programme.
Use the information in the passage and your own knowledge to answer the following
questions.

(a) People with AIDS die because they are unable to produce an immune response
to pathogens (lines 2-4).

Explain why this leads to death.

........................................................................................................................

........................................................................................................................

........................................................................................................................

........................................................................................................................

........................................................................................................................

........................................................................................................................

.......................................................................................................................

........................................................................................................................

........................................................................................................................(3)

(b) Explain why each of the following means that a vaccine might not be effective
against HIV.

(i) HIV rapidly enters host cells (lines 6-7).

...............................................................................................................

...............................................................................................................

...............................................................................................................

...............................................................................................................

............................................................................................................... (2)

(ii) HIV shows a lot of antigenic variability (lines 7-8).

...............................................................................................................

...............................................................................................................

...............................................................................................................

...............................................................................................................

...............................................................................................................(2)
(c) So far, these types of vaccine have not been considered safe to use in a mass
vaccination programme (lines 14-15).

Suggest why they have not been considered safe.

........................................................................................................................

........................................................................................................................

........................................................................................................................

........................................................................................................................

........................................................................................................................

........................................................................................................................

...................................................................................................

........................................................................................................................

........................................................................................................................
(3)
(Total 10 marks)
M1. (a) Virus/fungus/protozoan;
Neutral: named example
1

(b) Produces toxins;

Neutral: infects/colonises/invades cells

Damages cells/tissues/example given e.g. cell lysis;

2

(c) (i) (Antibodies) produced from a single clone of B cells/plasma cells;

Accept: hybridoma cell line instead of B cell/plasma
cell
Reject: idea that antibodies are cloned

OR

(Antibodies) produced from the same B cell/plasma cell;

1

(ii) (Specific) primary structure/order of amino acids;

(Specific) tertiary/3D structure;

(So) Only binds to/fits/complementary to one antigen;

Reject: active site for either point 2. or 3. only once
3

(d) (Rapid) treatment of carriers/infected cattle/disease;

Neutral: reference to rapid identification of infected
cattle

Can isolate/cull carriers/infected cattle;

Infected (dairy) products not sold/consumed/tracked;

Reduces spread of disease;

No need to kill/prevents the death of non-infected animals;

Neutral: ethical arguments
3 max
[10]

M2. (a) (i) Many people do not go to the doctor;

1

36000;
No marks awarded for working here as calculation
is very straightforward
1
 (b) Increases uptake of sodium ions/glucose/sugars / salts;

By co-transport channels/proteins;

Lowers water potential in cells/tissue;

Water moves out of intestine/into cells;

By osmosis;
Q Accept any correct alternative for lowering water
potential in cells.
3 max

(c) Same sugars/antigens on bacteria/nerve cells;

Do not accept references to same shape as
equivalent to complementary.

Bind with antibody/form antigen-antibody complex;

Reject react

Have complementary shape/fit binding site;

Reject active site
3

(d) Diaphragm will not move down/flatten /contract;

Ignore references to breathing out

Thoracic cavity/lung volume not increased so cannot breathe in;

2
[10]

M3.(a) 1. Microvilli;
1. Accept large surface area

2. Carrier proteins/co-transport proteins/membrane-bound

enzymes;

3. Many mitochondria;
Accept lots of ATP produced
2 max

(b) (i) Substance that causes an immune response/production of

antibodies;
Ignore foreign/non-self
1

(ii) 1. Not lipid soluble;

2. Too large (to diffuse through the membrane);

3. Antigens do not have the complementary shape / cannot

bind to receptor / channel / carrier proteins (in
membranes of other epithelial cells);
2 max
 (c) 1. (Vaccine contains) antigen / attenuated / dead pathogen;
1. Reject if in context of injection of vaccine

2. Microfold cells take up / bind and present / transport antigen (to

immune system / lymphocytes / T-cells);

3. T-cells activate B-cells;

3. Accept T-cells release cytokines

4. B-cells divide / form clone / undergo mitosis;

4. Accept plasma cells for B-cells

5. B-cells produce antibodies;

6. Memory cells produced;

6. Ignore T / B in reference to memory cells

7. More antibodies / antibodies produced faster in secondary

response / on reinfection;
7. Must be comparative
5 max
[10]

M4. (a) molecule (on cell surface);

that triggers immune response;
2

(b) (i) axes right way round and labelled;

2nd peak drawn higher;
steeper gradient on second rise;
3

(ii) because one dose does not give a high enough level of
antibody to be effective/ because the antibody falls after a while;
1

(iii) antigens are only single molecules/part of parasite;

do not actually cause disease;
2

(c) malaria sufferers would have parasites in red blood cells;

1
[9]

M8. (a) Girls are not sexually active/not likely to carry HPV/vaccine
may not work if already infected/few girls sexually active (at this age);
Neutral: girls are not sexually mature
Neutral: to provide better protection
Accept: provides immunity before sexually active
Neutral: girls are less likely to have it as could
mean the vaccine from the question stem
1
(b) Other (HPV) types have different antigens;

No memory cells for other types/memory cells not activated;

Antibodies cannot attach to antigen/correct antibodies not produced

/antibodies are not complementary;
Accept: refs. to antigenic variability
Accept: B cells for memory cells
Accept: memory cells cannot recognise antigen for
not activated
Accept: examples of memory cell activation
2 max

(c) More antigen;

More memory cells;

So more antibodies produced/antibodies produced quicker (if infected);

Accept: many/enough instead of more
Neutral: primary/secondary response
Accept: T cells/B cells/plasma cells instead of
antibodies
Reject: the idea that vaccines contain antibodies
Q Reject: antibodies fight/antibiotics
2 max

(d) Cancer takes years to develop/develops later in life;

Takes time for females to become sexually active/females must become

sexually active to obtain data;

Few people/only teenagers vaccinated;

Neutral: will take time to vaccinate 80% of young
girls
Accept: do not develop cancer instantly
2 max

(e) (Cervical cancer) can be caused by other types of HPV/other

factors/example given;

OR

(Some) women may have been infected (with HPV) before receiving
the vaccine;

OR

(As a precaution) in case vaccine does not work/a way of monitoring

if the vaccine has worked;
Accept: caused by other types of HPV in the
context of mutation
Neutral: to check for abnormal cells/that they are
immune to the virus
1
 (f) Virus cannot replicate/is destroyed/is not carried (in vaccinated people);

Non-vaccinated people more likely to contact vaccinated people;

Neutral: do not spread virus as in question stem
Must be in context of the individual and not the
population as in question stem
Q Do not allow disease is destroyed
Neutral: herd effect as given in the question stem
2
[10]

M11.(a) 1. Infected by / susceptible to (other) pathogen(s) / named disease caused

by a pathogen (from environment);
Context is where immune system cannot prevent or
stop these events
Allow attack / kill

2. Pathogen(s) reproduce / cause diease (in host);

MPs not given in context of HIV

3. Damage cells / tissues / organs;

4. Release toxins;
3 max

(b) (i) 1. (HIV enters cells) before antibodies can bind to / destroy it;
Ignore SAFETY comments
1. and 2. Relate to antibodies

2. Antibodies cannot enter cells (to destroy HIV) / stay in

blood;

OR

3. (Enters cells) before (secondary) immune response

caused / before memory cells have time to respond;
3. and 4. Relate to virus

4. So no antibodies present (to attack HIV);

OR

5. Vaccine taken up too quickly to cause immune response;

5. and 6. Relate to vaccine

6. So no antibodies / memory cells formed;

2 max

(ii) 1. Antigen (on HIV) changes;

 Accept mutates

2. (Specific) antibody / receptor no longer binds to (new)

antigen;
Ignore SAFETY comments

OR

3. Many different strains of HIV / many antigens present on

HIV;

4. Not possible to make a vaccine for all antigens / vaccine

may not stimulate an antibody for a particular antigen;
2 max

(c) 3 suitable suggestions;;;

QWC ignore reference to HIV cells

E.g.

1. Inactive virus may become active / viral transformation;

2. Attenuated virus might become harmful;

3. Non-pathogenic virus may mutate and harm cells;

4. Genetic information / protein (from HIV) may harm cells;

5. People (may) become / test HIV positive after vaccine used;

5. Vaccinated people may develop disease from
a different strain to that in the vaccine

6. This may affect their work/life;

6. May continue high risk activities and develop
or pass on HIV
3 max
[10]