Glycoproteins

• Glycoproteins are proteins that contain oligosaccharide

chains (glycans) covalently attached to their
polypeptide side-chains.

• They are one class of glycoconjugate or complex

carbohydrates—equivalent terms used to denote
molecules containing one or more carbohydrate chains
covalently linked to protein (to form glycoproteins or
proteoglycans) or lipid (to form glycolipids).

• The carbohydrate is attached to the protein in a

cotranslational or posttranslational modification in a
process is known as glycosylation.
• In proteins that have segments extending
extracellularly, the extracellular segments
are often glycosylated.

• Glycoproteins are often important

integral membrane proteins, where they
play a role in cell-cell interactions
N-linked protein glycosylation (N-glycosylation of N-glycans) at Asn residues
(Asn-x-Ser/Thr motifs) in glycoproteins.
 Some functions served by glycoproteins

Function Glycoproteins

Structural molecule Collagens, elastin, fibrins, bone matrix

Lubricant &protective agent Mucins

Transport molecule Transferrin, ceruloplasmin

Immunoglobins, histocompatibility antigens, compliment,
Immunologic molecule
interferons
Chorionoic gonadotropin, thyroid-stimulating hormone
Hormone
(TSH)
Enzyme Various, eg, alkaline phosphatase, proteases, glycosidases.

Cell attachment-recognition
cell-cell, virus-cell, bacterium-cell, and hormone receptors
site

Interact with specific

Lectins, selectins (cell adhesion lectins), antibodies
carbohydrates

Receptor Various proteins involved in hormone and drug action

Specific glycoproteins on the surface membranes of
Hemostasis & thrombosis
platelets
 N-glycosylation and O-glycosylation
• There are two types of glycosylation:
• In N-glycosylation, the addition of sugar chains can
happen at the amide nitrogen on the side chain of the
asparagine. Includes most circulating and membrane
bound glycoproteins

• In O-glycosylation, the addition of sugar chains can

happen on the hydroxyl oxygen on the side chain of
hydroxylysine, hydroxyproline, serine, or threonine.
Most are found in mucins though some are membrane
bound and others circulating.

• The sugar group(s) can assist in protein folding or

improve proteins' stability.
 Classes of glycoprotein
• (1) those containing an O-glycosidic linkage (ie, O-linked),
involving the hydroxyl side chain of serine or threonine and
a sugar such as N-cetylgalactosamine (GalNAc-Ser[Thr]);

• (2) those containing an N-glycosidic linkage (ie, N-linked),

involving the amide nitrogen of asparagine and N-
acetylglucosamine (GlcNAc-Asn); and

• (3) those linked to the carboxyl terminal amino acid of a

protein via a phosphoryl-ethanolamine moiety joined to an
oligosaccharide (glycan), which in turn is linked via
glucosamine to phosphatidylinositol (PI).
 Glycopeptide bonds
CH2 OH O NH2

H
O HN C CH2 CH COOH Asn
H
Glc OH H

OH O H

H HN C CH3 NAc
Type I N-Glycosyl linkage to Asn

CH2 OH NH2 CH2 OH NH2

O O CH2 CH COOH O O CH CH2

H H
H H
Glc OH H
Ser Glc OH H CH2 HOLys
OH O H OH H CH2

H HN C CH3 NAc H OH H2 N CH COOH

Type II O-Glycosyl linkage to Ser (Thr) Type III O-Glycosyl linkage to 5-HOLys
 Monosaccharides commonly found in eukaryotic
glycoproteins
The principal sugars found in human glycoproteins
Sugar Type Abbreviation
Galactose Hexose Gal
Glucose Hexose Glc
Mannose Hexose Man
N-Acetylneuraminic acid Sialic acid NeuAc
N-Acetylgalactosamine Deoxyhexose GalNAc
N-Acetylglucosamine Aminohexase GlaNac
Fucose Aminohexase Fuc
Xylose Pentose Xyl
 Functions of the oligosaccharide chains of glycoproteins

• Modulate physicochemical properties e.g solubility,

visicosity, charge & denaturation.

• Protect against proteolysis

• Affect proteolytic processing of precursor proteins to

smaller products
• Are involved in biologic activity e.g hCG

• Affect insertion into membranes, intracellular

migration & secretion

• Affect embryonic development & differentiation

• May affect sites of metastasis selected by cancer

cells.
 Mucins
• Mucins are glycoproteins with two major
characteristics:
a. a high content of O-linked oligosaccharides (the
carbohydrate content of mucins is generally more
than 50%); and

b. the presence of repeating amino acid sequences

(tandem repeats) in the center of their polypeptide
backbones, to which the O-glycan chains are
attached in clusters
• These sequences are rich in serine, threonine, and proline.
Although O-glycans predominate, mucins often contain a
number of N-glycan chains.

• Both secretory and membrane bound mucins occur.

• Secretory mucins are found in the mucus present in the

secretions of the gastrointestinal, respiratory, and
reproductive tracts. Mucus consists of about 94% water and
5% mucins, with the remainder being a mixture of various cell
molecules, electrolytes, and remnants of cells.

• Membrane-bound mucins participate in various cell-cell

interactions (eg, involving selectins;.
Properties of Mucins
 Proteoglycans

• These are proteins that contain covalently linked

glycosaminoglycans (GAG).

• The bound proteins are called core proteins while GAGs

are unbranched polysaccharide made of repeating
dissacharides, one component is always an amino sugar
(Glucosamine or galactosamine which or may not be
sullfated) & uronic acid (iduronate and glucuronate).
 EXAMPLES

• Dermatan sulfate is a glycosaminoglycan (formerly

called a mucopolysaccharide) found mostly in skin,
but also in blood vessels, heart valves, tendons, and
lungs.

• It is structurally similar to chondroitin sulfate

except for the 1,3 linkage of iduronic acid to
galactosamine instead of glucuronic acid.

• Dermatan sulfate may have roles in coagulation,

cardiovascular disease, carcinogenesis, infection,
wound repair, and fibrosis.
 Chondroitin sulphate
-1,3 -1,4
COO- CH2 OH

H O - O O
O OSO3
H H
OH H H

H H O H

H OH H HN C CH3

GlcUA GalNAc

GlcUA-Gal-Gal-Xyl-O-Ser link
Chondroitin sulfate Sulfate at 4 or 6 C of GalNAc
-1,3 glycosidic linkage
• Chondroitin sulfate is a sulfated glycosaminoglycan (GAG)
composed of a chain of alternating sugars (
N-acetylgalactosamine and glucuronic acid). It is usually
found attached to proteins as part of a proteoglycan.

• A chondroitin chain can have over 100 individual sugars,

each of which can be sulfated in variable positions and
quantities

• Chondroitin's functions largely depend on the properties of

the overall proteoglycan of which it is a part. These
functions can be broadly divided into structural as in bone,
cornea and cartilage and regulatory roles.

• However, this division is not absolute and some

proteoglycans have both structural and regulatory roles.
 Heparin
-1,4
COO- CH2 OSO3 -

O
-1,4
H H O H
H
H H
OH H OH H

O O

H OSO3 - H NHSO3-

GlcUA GlcNAc

GlcN and GlcUA or IdUA

Heparin N and O sulfate (C2,3,6)
-1,4 glycosidic linkage
> NAc
Heparan sulfate
< N and O sulfate
• Heparin, a highly-sulfated glycosaminoglycan, is
widely used as an injectable anticoagulant and has
the highest negative charge density of any known
biological molecule.

• Its blood anti-coagulation is achieved mostly by

endothelial cell-derived heparan sulfate
proteoglycans.

• Heparin is usually stored within the secretory

granules of mast cells and released only into the
vasculature at sites of tissue injury.
 Hyaluronate

COO- -1,3 CH2 OH -1,4

H O O O
OH
H H
OH H H

H OH O H

H OH H HN C CH3

GlcUA GlcNAc

No protein link
No sulfate
Hyaluronate
-1,3 glycosidic
• Hyaluronate (hyaluronic acid) is a non-sulfated
glycosaminoglycan distributed widely
throughout connective, epithelial, and neural
tissues.

• It is one of the chief components of the

extracellular matrix, contributes significantly to
cell proliferation and migration, and may also
be involved in the progression of some
malignant tumors
• Hyaluronate is a major component of the
synovial fluid where it is responsiblefor the
increase the viscosity of the fluid. Along with
lubricin it is one of the fluid's main lubricating
components.

• Hyaluronate is an important component of

articular cartilage, where it is present as a coat
around each cell

• Hyaluronan is also a major component of skin,

where it is involved in tissue repair
 Examples with fuctions

• Mucins, which are secreted in the mucus of the

respiratory and digestive tracts.

• They are O-linked glycoproteins with ser and thr as the

linking aa

• The sugars attached to mucins give them considerable

water-holding capacity and also make them resistant
to proteolysis by digestive enzymes.

• Glycoproteins are important for white blood cell

recognition, especially in mammals.
• Examples of glycoproteins in the immune system
are: molecules such as antibodies
(immunoglobulins), which interact directly with
antigens

• molecules of the major histocompatibility complex

(or MHC), which are expressed on the surface of
cells and interact with T cells as part of the adaptive
immune response.

• In cell attachment and recognition; They are

components of the zona pellucida, which surrounds
the oocyte, and is important for sperm-egg
interaction.
• They constitute the determinants of blood groups
ande the Rhesus factor. The ABO sustances are
glycosphingolipids but they present as glycoproteins
in secretions. They differ in structure by single sugars.

• Structural glycoproteins such as Collagens, which

occur in connective tissue. They help bind together
the fibers, cells, and ground substance of connective
tissue. They may also help components of the tissue
bind to inorganic substances, such as calcium in
bone.

• As transport molecules such as Transferrin (for iron),

ceruloplasmin (for copper)
• As hormones e.g Follicle-stimulating hormone,
Luteinizing hormone, Thyroid-stimulating hormone,
human chorionic gonadotropin, Alpha-fetoprotein,
Erythropoietin (EPO)

• As enzymes e.g alkaline phosphatase

• As receptors they are involved in hormone and drug

action.

• Important in Homeostasis (and thrombosis) on the

surface membranes of platelets

• Soluble glycoproteins often show a high viscosity, for

example, in egg white and blood plasma.
Differences between glycoproteins and proteoglycans:

1.Length of the chain is relatively short (usually 2-10

sugar residues) very long in GAGs.

2.Do not have repeating disaccharide units.

3.They are branched.

4.May or may not be negatively charged.

Fibrous portein

By
Dr. C. Nyamwange
 Fibrous proteins
• Fibrous proteins, also called scleroproteins, are one of the two
main classes of protein quaternary structure (the other being
globular proteins). Fibrous proteins are only found in animals.

• Fibrous proteins form long protein filaments, rod- or wire-like

shapes. They are usually inert structural or storage proteins.

• They are generally water-insoluble and are found as an

aggregate due to hydrophobic R groups that stick out of the
molecule. The amino acid sequences they are made from often
have limited residues with repeats.

• These can form unusual secondary structures, e.g. collagen triple

helix. The structures often contain 'cross-links' between chains,
for example cys-cys disulfide bonds between keratin chains.
 Main classes
• Keratins: found in hair, fingernails, and bird
feathers

• Collagens: the most abundant proteins in a

vertebrate body – found in connective tissues
such as cartilage

• Elastins: found in ligaments, around blood

vessels.
 Keratins
• Keratins are the main constituent of structures that grow
from the skin:

• the α-keratins in the hair (including wool), horns, nails, claws

and hooves of mammals. The -keratin helix is a right-
handed helix, the same helix found in many other
proteins.

• the harder β-keratins found in nails and in the scales and

claws of reptiles, their shells (chelonians, such as tortoise,
turtle, terrapin), and in the feathers, beaks, and claws of
birds

• The harder keratins are formed primarily in beta sheets.

However, beta sheets are also found in α-keratins.
• Keratins contain a high proportion of the smallest of the 20
amino acids, glycine, whose "side group" is a single
hydrogen atom; also the next smallest, alanine, with a small
and non-charged methyl group.

• In the case of β-sheets, this allows stearically-unhindered

hydrogen bonding between the amino and carboxyl groups of
peptide bonds on adjacent protein chains, facilitating their
close alignment and strong binding.

• Fibrous keratin molecules can twist around each other to

form helical intermediate filaments.
• In addition to intra- and intermolecular hydrogen bonds,
keratins have large amounts of the sulfur-containing amino
acid cysteine, required for the disulfide bridges that confer
additional strength and rigidity by permanent, thermally-
stable crosslinking—a role sulfur bridges also play in
vulcanized rubber.

• Human hair is approximately 14% cysteine. The pungent

smells of burning hair and rubber are due to the sulfur
compounds formed. Extensive disulfide bonding contributes
to the insolubility of keratins, except in dissociating or
reducing agents
• The more flexible and elastic keratins of hair have fewer
interchain disulfide bridges than the keratins in mammalian
fingernails, hooves and claws (homologous structures),
which are harder and more like their analogs in other
vertebrate classes.

• Hair and other α-keratins consist of α-helically-coiled single

protein strands (with regular intra-chain H-bonding), which
are then further twisted into superhelical ropes that may be
further coiled.

• The β-keratins of reptiles and birds have β-pleated sheets

twisted together, then stabilized and hardened by disulfide
bridges.
Structure of hair
• Hair -keratin is an elongated helix with somewhat thicker
elements near the amino and carboxyl termini.

•Pairs of these helices are interwound in a left-handed sense to

form two-chain coiled coils. These then combine in higher-
order structures called protofilaments and protofibrils.

•About four protofibrils—32 strands of -keratin altogether—

combine to form an intermediate filament. The individual two-
chain coiled coils in the various substructures also appear to be
interwound, but the handedness of the interwinding and other
structural details are unknown.
 Collagens
• Collagen is the main protein of connective tissue in
animals and the most abundant protein in mammals
making up about 25% of the whole-body protein
content.
• At least 19 distinct types of collagen made up of 30
distinct polypeptide chains (each encoded by a separate
gene) have been identified in human tissues. Although
several of these are present only in small proportions,
they may play important roles in determining physical
properties of specific tissues.
• Like the -keratins, collagen has evolved to provide strength. It is
found in connective tissue such as tendons, cartilage, the organic
matrix of bone, and the cornea of the eye.

• The collagen helix is a unique secondary structure quite distinct

from the helix. It is left-handed and has three amino acid residues
per turn

• Collagen is also a coiled coil, but one with distinct tertiary and
quaternary structures: three separate polypeptides, called  chains
(not to be confused with  helices), are supertwisted about each
other.

• The superhelical twisting is right-handed in collagen, opposite in

sense to the left-handed helix of the chains.
• The tropocollagen or "collagen molecule" is a subunit of
larger collagen aggregates such as fibrils.

• It is approximately 300 nm long and 1.5 nm in diameter,

made up of three polypeptide strands (called alpha
peptides), each possessing the conformation of a left-
handed helix (its name is not to be confused with the
commonly occurring alpha helix, a right handed
structure).

• These three left-handed helices are twisted together into

a right-handed coiled coil, a triple helix or "super helix",
a cooperative quaternary structure stabilized by
numerous hydrogen bonds.
• With type I collagen and possibly all fibrillar
collagens if not all collagens, each triple-helix
associates into a right-handed super-super-
coil that is referred to as the collagen
microfibril.
 Types of Collagen
• The collagen superfamily of proteins include more than twenty collagen types,as
well as additional proteins that have collagen like properties.
• The three polypeptide chains are held together by hydrogen bonds between the
chains.
• Variations in the aminoacid sequence of the α chains result in structural
components that are about the same size(approximately 100 aminoacids
long),but with slightly different properties.
• The α chains are combined to form various types of collagen found in the tissues.
• For example Type 1 collagen contains two α1 chains and one α2 chain (α1 ₂ α2 ).
• Type 2 collagen contains three α1(α1 ₃) chain.
• The collagens can be organized into three groups depending upon their location
and functions in the body.
(a) The chain of
collagen has a
repeating
secondary
structure unique
to this protein.

(c) Three
of these helices
wrap around
one another with a
right-handed twist.
Structure and composition of collagen

• A distinctive feature of collagen is the regular arrangement of

amino acids in each of the three chains of these collagen
subunits.

• The sequence often follows the pattern Gly-Pro-Y or Gly-X-

Hyp, where X and Y may be any of various other amino acid
residues. Vertebrate collagen typically contains about 35%
Gly, 11% Ala, and 21% Pro and 4-Hyp

• This kind of regular repetition and high glycine content is

found in only a few other fibrous proteins, such as silk fibroin.
75-80% of silk is (approximately) -Gly-Ala-Gly-Ala- with 10%
serine — and elastin is rich in glycine, proline, and alanine
(Ala), whose side group is a small, inert methyl group.
• Such high glycine and regular repetitions are never found
in globular proteins save for very short sections of their
sequence. Chemically-reactive side groups are not needed
in structural proteins as they are in enzymes and transport
proteins, however collagen is not quite just a structural
proteins.

• Only Gly residues can be accommodated at the very tight

junctions between the individual chains. The Pro and 4-
Hyp residues permit the sharp twisting of the collagen helix

• Proline and hydroxyproline confer rigidity on the collagen

molecule.
 Callagen formation
Inside the cell
• Three peptide chains are formed (2 alpha-1 and 1
alpha-2 chain) in ribosomes along the Rough
Endoplasmic Reticulum (RER). These peptide chains
(known as pre-pro-collagen) have registration
peptides on each end; and a signal peptide is also
attached to each

• Peptide chains are sent into the lumen of the RER

where the signal Peptides are cleaved inside the RER
to form pro-collagen chains
• Hydroxylation of lysine and proline amino acids occurs inside
the lumen. This process is dependent on Ascorbic Acid (Vitamin
C) as a cofactor
– Glycosylation of specific hydroxylated amino acid occurs
– Triple helical structure is formed inside the RER
– Procollagen is shipped to the golgi apparatus, where it is
packaged and secreted by exocytosis

Outside the cell

• Registration peptides are cleaved and tropo-collagen is formed
by procollagen peptidase.

• Multiple tropo-collagen molecules form collagen fibrils, and

multiple collagen fibrils form into collagen fibers

• Collagen is attached to cell membranes via several types of

protein, including fibronectin and integrin.
 Elastin
• Elastin is a connective tissue protein that is responsible
for properties of extensibility and elastic recoil in tissues.

• Although not as widespread as collagen, elastin is

present in large amounts, particularly in tissues that
require these physical properties, eg, lung, large arterial
blood vessels, and some elastic ligaments.

• Smaller quantities of elastin are also found in skin, ear

cartilage, and several other tissues.
• Elastin is a protein in connective tissue that is elastic and
allows many tissues in the body to resume their shape
after stretching or contracting.

• Elastin helps skin to return to its original position when it

is poked or pinched.

• Elastin is also an important load-bearing tissue in the

bodies of mammals and is used in places where
mechanical energy is required to be stored.

• Elastin is primarily composed of the amino acids glycine,

valine, alanine, and proline.
• It is a specialized protein with a molecular weight of 64 to 66
kDa, and an irregular or random coil conformation made up
of 830 amino acids.

• Elastin is made by linking many soluble tropo-elastin protein

molecules, in a reaction catalyzed by lysyl oxidase, to make a
massive insoluble, durable cross-linked array.

• The unique cross-links, called desmosine, made of four

Lysine connected through their side chains, which in turn
gives the elastin strong elastic properties

• Desmosine and isodesmosine are both found in elastin.

• Elastin serves an important function in arteries and is
particularly abundant in large elastic blood vessels such
as the aorta.

• Elastin is also very important in the lungs, elastic

ligaments, the skin, the bladder, elastic cartilage, and the
intervertebral disc above the sacroiliac.

• Tropoelastin is a water-soluble molecule with a

molecular weight of approximately 70,000 daltons.
Multiple tropoelastin molecules covalently bind together
with crosslinks to form the protein elastin that is very
prevalent in the body.