Chapter 5

Membrane Structure and Transport

Membranes
Separate the Cell from its environment w Separate the Cell into subcellular domains
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Different reactions can take place in the different domains

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Components
Phospholipids
Amphipathic - polar head group + hydrophobic tail

Cholesterol (4 ring structure)

Phospholipid Membranes
Phospholipids have a hydrophobic tail w This tail Can NOT hydrogen bond with water
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It is Non-polar It prefers the less polar environment of air or itself

Membrane Fluidity
Phospholipids keep their hydrophobic tails away from water -FLIP-FLOP is rare w Kinky Phospholipids with Unsaturated hydrocarbon tails are more mobile than Saturated ones w Cholesterol is GOOD for promoting membrane fluidity (at low temperatures) and stability (at high temperatures) w Without membrane fluidity, the enzymes wouldnt work, the cell would DIE
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Membrane Fluidity is due to the properties of the phospholipids and cholesterol

Fluid-mosaic model
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Mosaic of protein embedded in a sea of lipid

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Extracellular environment Carbohydrate Glycolipid Integral membrane protein Glycoprotein Phospholipid bilayer

Extracellular leaflet

HO

Polar

Cytosolic leaflet

Nonpolar

Peripheral membrane proteins

Cytosol

Cholesterol (found only in animal cells)

Polar

Membrane Proteins
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2 Main kinds
Integral - transmembrane, can NOT be removed by anything except detergent (removal of phospholipids) Peripheral - stuck to the outside or inside of the membrane by electrostatic (charged) interactions, can be removed by increased salt concentration

Most proteins are free to diffuse in the lipid bilayer w Unless they are attached to the cytoskeleton or extracellular matrix (ECM)
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Membrane Proteins
Can have different Structures w Can be modified by the addition of Carbohydrate
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Glycoproteins
Involved in cell-cell adhesion Cell-Cell recognition (immune system) Cell-Substrate recognition (binding to ECM) Cell protection - glycocalyx

Lipid
Targeting of protein to membrane

Membrane Proteins can MOVE!

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FRAP (fluorescence recovery after photobleaching )

Membrane proteins move laterally

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BUT
w 1070%

of membrane proteins cant/dont move (depends on cell type) w Association with cytoskelton w Association with extracellular matrix or other cells

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~25% of All Genes Encode Membrane Proteins

Membranes required for LIFE w 2030% of all genes are predicted to encode membrane proteins
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ALL domains- archaea, bacteria, and eukaryotes

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The functions of most/many membrane proteins are currently unknown

TEM of cell

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Freeze Fracture EM Information about protein structure

Membrane bilayer 0.1 m (a) Transmission electron microscopy (TEM)

Direction of fracture Transmembrane protein

Lipid bilayer (b) Freeze fracture electron microscopy (FFEM)

P face exposed

E face exposed

P face Cytosolic leaflet

E face Extracellular leaflet E face

P face 0.4 m

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So where do Eukaryotic membranes come from?

Cytosol and endomembrane system both needed w Lipid synthesis occurs at cytosolic leaflet of smooth ER w Fatty acid building blocks made by enzymes in cytosol or taken into cells from food
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P P

OH OH
O C C O

CoA CoA
O C C O P O

CH2 CH CH2 O C O C O Pi Cytosol

Choline head group

CH2 CH CH2
OH OH

2 fatty acids

2 activated molecules

Glycerolphosphate

Phosphatase Acyl transferase ER lumen Choline phosphotransferase Flippase

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Transfer of lipids to other membranes

Lipids in ER membrane can diffuse laterally to nuclear envelope w OR by vessicles to Golgi, lysosomes, vacuoles, or plasma membrane w OR.Lipid exchange proteins extract lipid from one membrane for insertion in another
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ER membrane proteins
Most transmembrane proteins synthesized on Rough ER membrane w From the Rough ER, membrane proteins can be transferred via vesicles to other regions of the cell
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Cytosol

Ribosome mRNA Signal peptidase

5 3

ER membrane ER lumen Channel NH3+ Cleaved ER signal sequence Transmembrane segment with 20 hydrophobic amino acids
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COO

NH3+

NH3+

Glycosylation
Carbohydrates attached to a lipid or protein w Protein glycosylation
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N-linked attachment of carbohydrate to nitrogen atom of asparagine side chain O-linked occurs only in Golgi, addition of sugars to oxygen atom of serine or threonine side chains

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mRNA

Ribosome 3

Dolichol lipid

Oligosaccharide transferase COO

P P Channel Amino acid sequence asparagineX-threonine or serine

NH3+

Carbohydrate tree attached to dolichol

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NH3+

Membranes - How they work..

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Selective permeability due to molecular organization of the membrane

Hydrophobic molecules can cross
steroids, hydrocarbons, carbon dioxide, oxygen

Very Small polar molecules can cross

water, ethanol

Larger polar molecules can NOT cross easily

amino acids, glucose,

Ions can NOT cross easily

H+, Na+, Ca+, K+, Cl-, etc.

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Cells maintain gradients

Living cells maintain a relatively constant internal environment different from their external environment w Transmembrane gradient
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Concentration of a solute is higher on one side of a membrane than the other

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Ion electrochemical gradient

Both an electrical gradient and chemical gradient

Glucose

Plasma membrane (a) Chemical gradient for glucosea higher glucose concentration outside the cell

+ + + + + + + + + + + + + + + +

+ + +

Cl Na+ K+

+ +

Plasma membrane
+ + +

(b) Electrochemical gradient for Na+more positive charges outside the cell and a higher Na+ concentration outside the cell

So how do Charged Molecules or Ions get into the Cell?

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Transport proteins
Integral membrane proteins Several different classes Some require energy (ATP) to work
(Active transporters)

Others do not require energy to work

(Passive transporters)

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ATP ADP + Pi

(a) Diffusionpassive transport

(b) Facilitated diffusionpassive transport

(c) Active transport

Passive Transport
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Transport DOWN a concentration gradient

Molecules move from a region of high concentration to a region of lower concentration This continues until there is an equal distribution Requires NO input of energy Due solely to diffusion, driven by kinetic energy of the molecules in solution

Passive transport continued

Passive transport does not require an input of energy w 2 types
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Passive diffusion
Diffusion of a solute through a membrane without transport protein

Facilitated diffusion
Diffusion of a solute through a membrane with the aid of a transport protein
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Tonicity
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Isotonic
Equal water and solute concentrations on either side of the membrane

Hypertonic
Solute concentration is higher (and water concentration lower) on one side of the membrane

Hypotonic
Solute concentration is lower (and water concentration higher) on one side of the membrane
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Outside the cell

Inside the cell

The solution and cell are isotonic

Isotonic

Hypertonic

The solution is hypertonic to the cell

The solution is hypotonic to the cell

Hypotonic
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Osmosis
Water diffuses through a membrane from an area with more water to an area with less water w If the solutes cannot move, water movement can make the cell shrink or swell as water leaves or enters the cell w Osmotic pressure- the tendency for water to move into any cell
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Osmosis - Movement of Water to equalize solute concentration

Animal cells must balance extracellular and intracellular solute concentrations to keep size and shape w Crenationshrinking in a hypertonic solution
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A cell wall prevents major changes in cell size w Turgor pressurepushes plasma membrane against cell wall Maintains shape and size w Plasmolysisplants wilt because water leaves plant cells
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Agre Discovered That Osmosis Occurs More Quickly in Cells with Transport Proteins That Allow the Facilitated Diffusion of Water
Water passively diffuses across plasma membranes w But sometimes water to moves across the plasma membrane at a much faster than predicted by passive diffusion w Protein that was abundant in red blood cells and kidney cells, but not found in many other cell types
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CHIP28 Striking difference was observed between frog oocytes that expressed CHIP28 versus the control
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Aquaporins Agre was awarded the Nobel Prize in 2003 for this work

Transport proteins
Transport proteins enable biological membranes to be selectively permeable w 2 classes
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Channels Transporters

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Channels
Form an open passageway for the direct diffusion of ions or molecules across the membrane w Aquaporins w FAST
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Most are gatedopen or close

Ligand-gated Intracellular regulatory proteins Phosphorylation Voltage-gated Mechanosensitive channels (dancing ear cell movie)

Gate opened Gate closed

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Transporters
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Also known as carriers Conformational change transports solute Main way for cell to move organic molecules, such as sugars, amino acids, and nucleotides Key role in export Slower than Channels

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Transporter types
Uniporter
single molecule or ion

Symporter/ cotransporter
2 or more ions or molecules transported in same direction

Antiporter
2 or more ions or molecules transported in opposite directions

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Active transport
Movement of a solute across a membrane against its gradient from a region of low concentration to higher concentration w Energetically unfavorable and requires the input of energy w Primary active transport
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Directly use energy to transport solute

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Secondary active transport

Use pre-existing gradient to drive transport of solute
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Active Transport
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Transport UP a concentration gradient

Molecules move from a region of low concentration to a region of higher concentration This continues until the cell runs out of energy! Requires an input of energy

Driven by the hydrolysis of ATP or sometimes a Proton (H+) gradient (Secondary

active transport)

Pump
conformational changes to an energy source, such as ATP-driven pumps
ATP hydrolysis can be uniporters, symporters, or antiporters Active transport
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ATP Ion Pumps Generate Ion Electrochemical Gradients

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Na+/K+-ATPase
Actively transport Na+ and K+ against their gradients by using the energy from ATP hydrolysis 3 Na+ exported for 2 K+ imported into cell
Antiporter Electrogenic pump- export 1 net positive charge

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Exocytosis/ Endocytosis
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Transport larger molecules and particles

proteins , polysaccharides

Exocytosis
Transport of material in vesicles dumped outside

Endocytosis
transport of materials into cell in newly formed vesicles made from plasma membrane Receptor-mediated endocytosis Pinocytosis Phagocytosis
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Cellular Membranes have a polarity or sidedness

The inside differs from the outside

Exocytosis

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Receptor Mediated Endocytosis

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Sites of Receptor Mediated Endocytosis

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Honeycomb structures called clathrin cages Note other smooth vesicles probably involved in fluid phase pinocytosis