GKM/NSB113/BLD.BDFL/MYO.

HAE/2018
MSB 103: HAEMOPOETIC SYSTEM AND BODY FLUIDS

~ Glycosaminoglycans (Proteoglycans)
~ Glycoproteins
~ Mucins
LECTURER:
DR. GEOFFREY K. MAIYOH
Department of Medical Biochemistry

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What do they have in common?
• They are associations between protein and
carbohydrates (however,……different kinds
and in different proportions)

Remember ?
• Carbohydrates – polymers of simple sugars
(monosaccharide)
• Proteins – polymers of amino acids
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 Carbohydrates - structure

Cn (H)2nOn

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 Protein - structure

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Why are they of interest?

• Have been shown to perform diverse functions

• Have been discovered to be abundant in living
organisms.
• They appear in nearly every biological process studied.

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Functions
• Many glycoproteins have structural functions e.g. in certain bacteria
the slime layer that surrounds the outermost components of cell walls
are made up of glycoproteins of high molecular weight.
• Glycoproteins also form connective tissues such as collagen.
• They are also found abundantly in the blood plasma where they serve
many functions e.g. Prothrombin, thrombin, and fibrinogen are all
glycoproteins that play an intricate role in the blood clotting
mechanism
• Mucin glycoproteins are also used as lubricants.

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• Protection: High molecular weight polymers called mucins are found on
internal epithelial surfaces.
• They form a highly viscous gel that protects epithelium form chemical,
physical, and microbial disturbances e.g. the human digestive tract,
urinary tract, and respiratory tracts.
• Reproduction: Glycoproteins found on the surface of spermatozoa
appear to increase a sperm cell's attraction for the egg by altering the
electrophoretic mobility of the plasma membrane.
• Actual binding of the sperm cell to the egg is mediated by
glycoproteins serving as receptors on the surface of each the two
membranes.
• "Cervical mucin" is a glycoprotein found in the cervix of animals that
regulates access of spermatozoa to the upper reproductive tract.
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Other functions
• Cell-cell adhesion and cell-cell interactiions
• Hormones – many e.g. HCG and erythropoietin
• Enzymes: three types i.e. oxidoreductases, transferases, and hydrolases
• Carriers: e.g. bind and transport vitamins, hormones, cations, and
other substances.
• Vision: forms the outer membranes of retinal rods

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More roles
• Immunological: The interaction of blood group substances with
antibodies is determined by the glycoproteins on erythrocytes.
• Adding or removing just one monosaccharide from a blood group
structure, the antigenicity and therefore a person's blood type can be
• B and T cells contain surface glycoproteins that attract bacteria and
bind them
• Protein targeting: Enable protein to reach its ultimate
destination in the cell or organism.

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 Glycoproteins
Mediate Cell Recognition

Your ABO blood type is determined by what sugars

you have in a particular oligosaccharide side chain
on one of the proteins that lies on the surface of
your red blood cells

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Why they are versatile
• As a direct result of their structure Proteins weds Carbs

i.e.
• Are composed of a peptide
chain joined to carbohydrate
moieties.
• Peptides and carbs. can perform
numerous roles independently

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The glycopeptide bonds
• There are two broad categories of structures. (fig. on next
slide)

1. The carbohydrates are either linked N-glycosidically

or

2. O-glycosidically to their constituent protein/peptides.

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glycopeptide bond

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Type I N-Glycosyl linkage to Asn

CH2 OH O NH2
Glycopeptide
H
H
O HN C CH2 CH COOH Asn bonds
Glc OH H

OH O H

H HN C CH3
NAc
Within these broader categories, there can be
Type II O-Glycosyl linkage to Ser (Thr) fine structural differences which account for the
CH2 OH NH2
large diversity of functions
O O CH2 CH COOH
H
H

Glc OH H Ser
OH O H Asn: asparagine Glc: glucose
NAc: N-acetyl Ser: serine
H HN C CH3
NAc Thr: threonine HOLys: 5-hydroxylysine

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Glycosaminoglycans
• Glycosaminoglycan (GAGs) are large complexes of negatively
charged heteropolysaccharide chains. They are generally
associated with a small amount of protein, forming proteoglycans,
which typically consist of > 95% CHO.

• Note: this is in comparison to the glycoproteins, which consist

primarily of protein with a small amount of CHO

CHO = Carbohydrate

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GAGs and Water
• Glycosaminoglycans have the special ability to bind large
amounts of water, thereby producing the gel-like matrix
that forms the basis of the body’s ground substance

• The viscous, lubricating properties of mucous secretions also

result from the presence of GAGs, which led to the original
naming of these cpds as mucopolysaccharides

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Structure of GAGs
• GAGs are long, unbranched, heteropolysaccharide chains
generally composed of a repeating disaccharide unit [acidic
sugar-amino sugar]n
• The amino sugar is either;
1. D-glucosamine or
2. D-galactosamine,

The amino group is usually acetylated, thus eliminating its

+ve charge.
• The amino sugar may also be sulfated on Carbon atom no. 4 or
6 or on a non-acetylated nitrogen
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Figure 14.1. Repeating disaccharide unit.

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• The acidic sugar is either;

1. D-glucuronic acid or
2. its C-5 epimer, L-iduronic acid (fig. on next slide)

• These acidic sugars contain carboxyl groups that are

negatively charged at physiologic pH &, together with the
sulfate groups, give GAGs their strongly –ve nature

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Some monosaccharide units found in glycosaminoglycans

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Relationship between GAG structure and function
- Because of their large # of –ve charges, these heteropolysaccharide
chains tend to be extended in solutions.
- They repel each other and are surrounded by a shell of water molecules.

- When brought together, they “slip” past each other, much as two
magnets with the same polarity seem to slip past each other.

- This produces “slippery” consistency of mucous secretions & synovial

fluid

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GAGs and resilience
- When a soln of GAGs is compressed, the water is “squeezed out” &
the GAGs are forced to occupy a smaller volume.

- When the compression is released, the GAGs spring back to their

original, hydrated volume because of repulsion of their –ve charges.

- This property contributes to the resilience of synovial fluid & the

vitreous humor of the eye.

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 Figure 14.3. Resilience of glycosaminoglycans.
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B. Classification of GAGs
- The six major classes of GAGs are based on;
i. Monomeric composition
ii. Type of glycosidic linkages
iii. Degree & location of sulfate units
- The structure of the GAGs & their distribution in the body
is illustrated in the Next Slide.

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GAGs

1. Chodroitin 4- and 6- sulphate

2. Keratan sulfates
3. Hyaluronic acid
4. Dermatan sulfate
5. Heparin
6. Heparin sulfate

Know

• Monomers
• Where found
• Function

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Symbols for monomeric units
• Gal – Galactose
• GalN - Galactosamine
• GalNAC – N-acetylgalactosamine
• Glc – Glucose
• GlcN – Glucosamine
• GlcNAC – N-acetylglucosamine
• GlcUA– Glucuronic Acid
• IdUA – Iuduronic Acid

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Chondroitin -4 and 6- Sulfate

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Keratan Sulfates I and II

Acetyl group = -COCH3

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Hyaluronic Acid

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Structure of GAGs – proteoglycan
- All of the GAGs, except hyaluronic cid, are found covalently attached to
proteins, forming proteoglycan monomers. • Monomer
• Aggregate
Structure of proteoglycan monomer:
- A proteoglycan monomer consists of a core protein to which the linear GAG
chains are covalently attached
- These chains which may each be composed of >100 monosacch, extend
out from the core protein & remain separated from each other because of
charge repulsion.

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-The resulting structure resembles a “bottle brush”
-In cartilage proteoglycan, the species of GAGs include chondroitin
sulfate & keratan sulfate

"Bottle-brush" model of a cartilage proteoglycan monomer.

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Linkage between the carbohydrate chain & the protein
- This linkage is most commonly through a trihexoside (galactose-galactose-xylose) & a
ser residue, respectively. An O-glycosidic bond is formed b/w the xylose & the hydroxyl
group of the ser.

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Proteoglycan aggregates
- The proteoglycan monomers associate with a molecule of
hyaluronic acid to form proteoglycan aggregates.
- The association is not covalent, but occurs primarily
through ionic interactions b/w core protein & the
hyaluronic acid
- The association is stabilized by additional small proteins
called link proteins

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Proteoglycan aggregate.

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Glycoproteins
- Glycoproteins are proteins to which oligosacch’s are covalently
attached.
- They differ from proteoglycans in that length of glycoproteins’
CHO chain is relatively short (usually 2-10 sugar residues in
length, although they can be longer), whereas it can be very long
in the GAGs.
- In addition, whereas GAGs have diglycosyl repeat units, the
CHO’s of glycoproteins do not have serial repeats.
- The glycoprotein CHO chains are often branched instead of
linear, & may or may not be negatively charged.
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Structure of glycoprotein oligosaccharides

- The oligosaccharide components of glycoproteins are generally branched

heteropolymers composed primarily of D-hexoses.

Structure of the linkage between carbohydrate and protein

- A glycoprotein may contain only one type of glycosidic linkage (N- or O-
linked), or may have both O- and N-linked oligosacchs within same
molecule
- For N-linked, the sugar chain is attached to the amide group of an
asparagine side chain, while for O-linked, to a hydroxyl group of either ser
or thr R-group.

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N- and O-linked oligosaccharides

O-linked oligosaccharides:
- The O-linked oligosaccharides may have one or more of a
wide variety of sugars arranged in either a linear or branched
pattern.

- Many O-linked oligosaccharides are found as membrane

glycoprotein components or in extracellular glycoproteins.
- E.g., O-linked oligosaccharides help provide the ABO blood group
determinants

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N-linked oligosaccharides:
• The N-linked oligosaccharides fall into 2 broad classes:
i. complex oligosaccharides
ii. high-mannose oligosaccharides. (Next slide)

• Both contain same core pentasaccharide , but the complex

oligosacch’s contain a diverse group of additional sugars, e.g.,
N-acetylglucosamine (GlcNAc), L-fucose (Fuc), N-
acetylneuraminic acid (NANA), whereas the high-mannose
oligosacch’s contain primarily mannose (Man)

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Complex (top) and high-mannose (bottom)
oligosaccharides.

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Mucins
• Are major components of the extracellular mucus
blanket.
• They are high-molecular-mass glycoconjugates with
hundreds of oligosaccharide chains in O-glycosidic
linkages to a protein backbone.
• They protect and lubricate mammalian epithelia

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Mucin Structure
• The CHO component includes a six carbon sugar with a carboxyl
group as a functional group (see figure below)

This is called sialate, and whenever its part of a OH

HO OO O
structure, has negative charge
OH C
Ac N
Mucin contains lots of sugars, with very short side OH
chain, only two monosaccharides long.
OH
sialate
(N-acetylneuraminate)

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Mucin structure
• Mucin give mucous their main property, viscous property
of saliva and mucous secretions in general.
• The sialate and to a lesser extent the other monosaccharide
are very hydrophilic and thus bind lots of water.
• They form very extended structures because of the negative
charge.

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Mucins: salivary glycoproteins

OH
~ ~
HO OO O–
OH C galNAc
Ac N 2 sialate
OH
OH • Have approximately 800 short (disaccharide) side
chains
sialate
(N-acetylneuraminate) • Terminal sugar is sialate
• anionic sugar
• Very hydrophilic, extended structure

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Mucins: modification &
aggregation
• The enzyme sialidase
(neuraminidase) catalyzes hydrolysis
of sialates from mucins ~ ~
• It is secreted by oral bacteria
• products: x H 2O galNAc
sialidase sialate
• Are less hydrophilic, less H2O-soluble, x
more folded, more aggregated

• This forms part of the enamel pellicle ~

& dental plaque matrix ~

~
~
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Dental Plaques
• Part of that meshwork is contributory to dental problems.

• This happens as a consequence of changing the mucins from

their freshly secreted form that is water soluble, to a less
soluble form that allows it to aggregate and bind to surfaces
in general.

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Role of glyco moiety in controlling protein lifetime

• Many blood proteins have a glyco chains with a terminal sialate

• Endothelial surface sialidases slowly remove sialates from these circulating
proteins
• The rate of sialate removal depends on protein's structure
sialoglycoprotein: sia–gal–glcNAc–[core sugars]–protein

asialoglycoprotein: gal–glcNAc–[core sugars]–protein

• Now-exposed gal–glcNAc… residues bind to asialoglycoprotein receptor
on liver cell surface
• The protein is then endocytosed & broken down

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21
Summary – For your refreshment
• GAGs are long, negatively charged, unbranched heteropolysacch chains generally
composed of a repeating disaccharide unit [acidic sugar-amino sugar]n
• The amino sugar is either D-glucoamine or D-galactosamine in which the amino
group is usually acetylated, thus eliminating its +ve charge
• The amino sugar may also be sulfated on C-4 or 6 or on a non-acetylated nitrogen.
• The acidic sugar is either D-glucuronic acid or its C-5 epimer, L-iduronic acid.
These cpds bind large amounts of water, thereby producing the gel-like matrix
that forms the basis of the body’s ground substance
• The viscous, lubricating properties of mucous secretions are also caused by the
presence of GAGs which led to the original naming of these cpds as
mucopolysaccharides
• As essential components of cell surfaces, GAGs play an important role in
mediating cell-cell signaling & adhesion.
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• There are 6 major classes of GAGs, including chondroitin 4- & 6-sulfates, keratan
sulfate, dermatan sulfate, heparin, heparan sulfate & hyaluronic acid.
• All of the GAGs, except hyaluronic acid, are found covalently attached to protein,
forming proteoglycan monomers, which consist of a core protein to which the
linear GAG chains are covalently attached.
• The proteoglycan monomers associate with a molecule of hyaluronic acid to form
proteoglycan aggregates
• GAGs are synthesized in the ER & Golgi. The polysacch chains are elongated by
sequential addition of alternating acidic & amino sugars, donated by their UDP-
derivatives. The last step in synthesis is the sulfation of some of the amino sugars.
The source of the sulfate is 3’-phosphoadenosyl-5`-phosphosulfate.

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 Thanks for your attention

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