Module BIO00004C Molecular Biology and Biochemistry Lectures 16-17 Christoph Baumann (christoph.baumann@york.ac.

uk)

Introduction to Structure and Function of Carbohydrates

Learning Outcomes
At the end of these lectures you should know: what are carbohydrates the basic structure of some important monosaccharides, disaccharides and polysaccharides the functions of carbohydrates in prokaryotic and eukaryotic cells

Starch Cellulose

Sucrose Chitin

Extensive roles in all forms of life As monosaccharides and disaccharides they are.. used as energy stores, fuels and metabolic intermediates important in the structural backbone of DNA and RNA As polysaccharides they are.. structural elements
walls of plants and bacteria extracellular matrix connective tissue lubrication of skeletal joints

linked to proteins and lipids

cell-cell recognition cell adhesion intracellular localisation tags on glycoproteins determine metabolic fate of glycoconjugates

Carbohydrates in Biomedicine
Erythropoietin glycosylated protein in serum that stimulates red blood cell
production cells

A, B and O blood groups 3 different carbohydrates on surface of blood Heparin glycosaminoglycan commonly used as a blood thinner Arthritis proteolytic degradation of collagen in cartilage Congenital lysosomal storage disorders (mucolipidosis II) enzymes
responsible for degrading glycosaminoglycans are missing from lysosomes surface glycoproteins (hemagglutinin)

Influenza virus enters cells by binding to sialic acid residues present on cell-

Carbohydrates in Biotechnology
Agarose (crosslinked D-galactose and 3,6-anhydrogalactose) Dextran (crosslinked !-1,6-D-glucose) Lectins applications as anti-tumor and anti-infective compounds Glycodendrimers DNA- or drug-delivery systems, materials science

Glyceraldehyde, the simplest carbohydrate

(Fischer projection) H C H OH H H C C H D-glyceraldehyde H C HO C H H H C C OH enantiomers O OH OH H C HO H L-glyceraldehyde O C C H O O

mirror image

CH2OH

CH2OH

How to Visualise the Fischer Projection of an Organic Molecule?

D-Aldoses containing three, four, five and six carbons

Triose

(not mirror images)

Diastereomers

Tetrose

Pentose

Hexose

Epimers

D-Ketoses containing three, four, five and six carbons

Tetrose

Diastereomers

Pentose

Hexose

Monosaccharides of the same chemical composition can vary tremendously in their structure For example: Hexose C6H12O6 48 possible structures Aldoses: 8 D, 8 L, ! 8 D, 8 L, " Ketoses: 4 D, 4 L, ! 4 D, 4 L, " 32 16

Ring formation
30 %
H
1C

CH2OH H OH

O
6

H
4

O
H
2

H
1

H HO H H

2 3 4 5 6

C C C C

OH H OH OH H
4

CH2OH H OH

H H
1 2

HO

OH

O
H OH

H O

OH

!-D-Glucose
(!-D-Glucopyranose)

HO

CH2OH

CH2OH H OH

D-Glucose

(open chain) <1%

hemiacetal

70 %
4

H HO

O
H
2

OH
1

OH

"-D-Glucose
("-D-Glucopyranose)

Reducing Sugars
Fehlings reagent

"-D-glucopyranose Glucose (open chain) Oxidised open chain

Aldehyde group is easily oxidised by Cu2+ (Fehlings reagent) thus they are known as reducing sugars

Different ways to depict "-D-Glucose

H
1C

O
6

H HO H H

2 3 4 5 6

C C C C

OH H OH OH
4

CH2OH H OH O H
2

H HO

OH
1

H HO 4 HO

CH2OH H
2

O
1

5 H 3

OH

OH

OH

CH2OH

Haworth

Fischer

Conformation : chair / all equatorial

Space-filling model

5 4 3

1 2

Chair and Boat Forms of !-D-Glucopyranose

a e
HO

a
CH2OH H O

e
OH

HO

CH2OH O H

HO H

HO

chair form

OH

HO H

boat form

OH

a = axial position

e = equatorial position

Fructose forms pyranose and furanose rings

H H
5 1 CH2OH 2 6

H H OH

O
HO
3

CH2OH
2

H H
5 6

HO 4 H H

H H OH

O
HO
3

CH2OH
2

HO 4 H

OH

C C C C

O H OH OH
6

HO H H

!-D-Fructopyranose
HOCH2
5

3 4 5 6

O
H HO
3

CH2OH
2

HOCH2
5

O
H HO H

CH2OH
2 3 OH

CH2OH

H 4 OH H

H 4 OH

D-Fructose

(open chain)

!-D-Fructofuranose
6

HOCH2
5

O
HO H

OH
2 3 CH2OH 1

H H 4 OH

hemiketal

"-D-Fructofuranose

Ribose
5

HOCH2
4

O H OH

OH
1 2 H

H H 3 OH

"-D-Ribose
5

HOCH2
4

O H

OH
1 2 H

H H 3 OH

H H

2-Deoxy-"-D-Ribose

2-deoxy

Glycosidic linkage
6

CH2OH H OH H O H
2

CH2OH H OH H O H
2

HO
4

OH
1 4

OH
1

H 3

HO 3

OH

OH

"-D-Galactose
H2 O
6

"-D-Glucose
6

CH2OH H OH H O H
2 1

CH2OH H OH O H
2

HO
4

OH
1

4 3

H 3

OH

OH

Gal-"-1,4-Glc

Lactose

CH2OH H OH O H
2

H
4

H
1

HO
6

CH2OH H OH H O H
2

H CH2OH H OH O H
2 6

OH

O O
H HO H
2 3 CH2OH 1

H
4

H
1

H
1

HOCH2
5

HO 3

4 3

OH

H 4 OH

OH

OH

Glc-!-1,4-Glc

!- D-Glc-1,2-"-D-Fru

Maltose

Sucrose

Some important sugar derivatives

6

CH2OH H OH O H
2

H
4

OH
1 6

HO

H H
4

CH2OH H OH O H
2

CH2OH H O H O H
2

NH2

OH
1 4

OH
1

D-Glucosamine (GlcN) O C6
H
4 5

HO

HO 3

NH C CH3 O

NH C CH3 O

O
O H
2

H OH H

OH
1

HO 3

N-Acetyl-D-Glucosamine (GlcNAc)

H3 C

C H

COOH

OH

D-Glucuronic acid (GlcUA)

N-Acetylmuramic acid (NAM)

Phosphorylated sugars are key intermediates in energy generation and biogenesis

CH2OPO32H OH H O H H
1

H
4

CH2OPO32C O CH2OH

CHO H C OH CH2OPO32-

HO 3

OH 2 OH

Glucose 6-phosphate (G-6P)

Dihydroxyacetone phosphate (DHAP)

Glyceraldehyde 3-phosphate (GAP)

Polysaccharides
Homopolysaccharides
Unbranched Branched

Heteropolysaccharides
Two monomer types, unbranched Multiple monomer types, branched

Polysaccharides for the storage of glucose

Storage of glucose: Glycogen

--Glc-!-1,4-Glc-!-1,6 branching every 8-12 residues

O
O

Glycogen granules in liver cell

O O O O O

non-reducing ends O
O O

" !

O O

reducing end

Storage of glucose in plants: Starch

Starch granules in plant cell

!-1,4 linkage

Amylose 1) !-Amylose --Glc-!-1,4-Glc-2) Amylopectin --Glc-!-1,4-Glc-!-1,6-branching every 24 - 30 residues

Nonreducing ends Reducing ends

Amylopectin

Polysaccharides: Structural roles

Plant cell walls

Cellulose: --Glc-"-1,4-Glc--

Primary wall Plasma membrane

Pectin Cellulose Hemicellulose

Microfibril

Storage versus structural polysaccharides

CH2OH H O H OH H O H H OH O H CH2OH H OH H O H H OH O H CH2OH H OH H O H H OH O H CH2OH H OH H O H H OH O H CH2OH H OH H O H H OH O H CH2OH H OH H O H H OH O

!-Amylose --Glc-!-1,4-Glc--

CH2OH O H H OH H O H OH O H H

CH2OH H OH H O H OH O H H

CH2OH H OH H O H OH O H H

CH2OH H OH H O H OH O H H

CH2OH H OH H O H OH O H H

CH2OH H OH H O H OH O H

Cellulose --Glc-"-1,4-Glc--

CH2OH O H H OH H O H NH C O C CH3 O H

CH2OH H OH H O H NH C O C CH3 O H

CH2OH H OH H O H NH C O C CH3 O H

CH2OH H OH H O H NH C O C CH3 O H

CH2OH H OH H O H NH C O C CH3 O H

CH2OH H OH H O H NH C O C CH3 O H

Chitin --GlcNAc-"-1,4-GlcNAc-

Peptidoglycan in bacterial cell envelope

--MurNAc-"-1,4-GlcNAc-6

CH2OH

CH2OH H OH O H
2

O
3

O H
2 1

4 3

N-Acetyl-D-glucosamine (GlcNAc)
O

H3 C

lactyl
O

CH H C O

NH2 C CH3 O

NH2 C CH3
M M M G G G M M M G G G M

Lysozyme
M M G G G

N-Acetylmuramic acid (MurNAc or NAM)

oligopeptide

Penicillins inhibit glycopeptide transpeptidase activity

G M
L-Ala D-Glu DAP D-Ala D-Ala D-Ala D-Ala DAP D-Glu
O

M
L-Ala

D-Ala

D-Glu DAP D-Ala D-Ala D-Ala DAP D-Glu L-Ala

L-Ala

H3N

CH CH2 CH2

M
C O

CH2 O HC NH3

DAP = diaminopimelic acid

Penicillins inhibit glycopeptide transpeptidase activity

G M
L-Ala D-Glu DAP D-Ala D-Ala D-Ala D-Ala DAP D-Glu L-Ala D-Ala

M
L-Ala D-Glu DAP D-Ala D-Ala

D-Ala DAP D-Glu L-Ala

M
R H N O N O C O CH3 O S CH3

"-lactam ring

Bacterial cell envelope / Peptidoglycan

Gram positive Staphylococcus aureus
Peptidoglycan Polysaccharide (cell wall) coat Lipoteichoic acid MurNAc GlcNAc MurNAc

Lipid bilayer membrane

Tetrapeptide

(Gly)5

Gram negative Escherichia coli

Lipopolysaccharide Outer lipid membrane Periplasmic space MurNAc GlcNAc MurNAc

Peptidoglycan

Lipid bilayer membrane

Cross-linked Tetrapeptides

Roles of carbohydrates in cellular recognition processes

Glycoproteins: glycosidic bonds between proteins and carbohydrates

N-linked GlcNAc

O-linked GalNAc

Glycoproteins
Example: N-Glycans
Man
O O

Primary
Glc
O

Man Man Man

O O O O O

Man Man

Man

O O

NH Asn

Man

GlcNAc

Glc

Glc

Man

Man Man Man Man

O O O O

O O

Man Man

Man

O O

NH Asn

Man

GlcNAc

Processing

Man

Glycosylation in the Cell

plasma membrane

Different glycosylation enzymes are present in each compartment (cisternae) of the Golgi

trans
Golgi

cis

ER ER

N-linked glycosylation begins in the endoplasmic reticulum (ER) and continues in the Golgi

O-linked glycosylation occurs in the Golgi

Processing

Man Man

Man

NH Asn

GlcNAc

Sia

Modification
O O

Man Gal
O O

L-Fuc O
O O O

Man
O

O O

Man

NH Asn

Sia

Gal

O O

GlcNAc

Carbohydrates as components of the extracellular matrix

Extracellular Matrix (ECM)

Actin filaments Integrin

Proteoglycan
Cell adhesion protein
PM

Fibronectin

Multiadhesive protein Collagen fiber

Collagen

PM

Plasma membrane

Glycosaminoglycans: Components of the extracellular matrix

COO COO COO
O O O O O O

Hyaluronic acid

"-1,3 GlcNAc GlcUA

"-1,4

OSO3 OSO3 OSO3 COO

O O

COO
O

COO
O

Chondroitin-6-sulfate

"-1,3 GalNAc GlcUA -6-sulfate OSO3 OSO3 Gal

O

"-1,4

OSO3 Gal
O O

Keratan sulfate

Gal

"-1,4

"-1,3

GalNAc -6-sulfate

Proteoglycan structure in cartilage

Hyaluronate Chondroitin sulfate

Proteoglycan

Core protein

Keratan sulfate

Link protein