EXTRACELLULAR

MATRIX
Mrs. OFELIA SOLANO SALUDAR
Department of Natural Sciences
University of St. La Salle
Bacolod City

 Many animal cells are intrinsically linked to other

cells and to the extracellular matrix (ECM).
Cell surface molecules bind to other cells, or to
other components of the ECM. They also play a
role in mutual recognition of similar cell types.
Bone and cartilage are mostly ECM plus a very few
cells. Connective tissue that surrounds glands and
blood vessels, is a gelatinous matrix containing
many fibroblast cells.

The ECM contains 3 classes of molecules:

structural proteins (collagens and elastins)
protein-polysaccharide complexes to embed the
structural
proteins
(proteoglycans)
adhesive
glycoproteins
to attach cells
to matrix
(fibronectins
and laminins).

PROTEOGLYCANS
1.PROTEOGLYCANS are composed of a core protein to which
glycosaminolycans (GAGs) are attached. GAGs consist of
repeating disaccharide subunits.
One of the two sugars in the disaccharide is often an amino
sugar (N-acetyl-glucosamine or N-acetyl-galactosamine; usually
with an attached sulfate group) and the other is a sugar or sugar
acid (galactose or glucuronate).
Chondroitin sulfate, keratan sulfate, heparan sulfate and
hyaluronate are the most common GAGs.

Each of the four classes of GAGs is

formed by polymerization of monomer
units into repeats of a particular
disaccharide and subsequent
modifications, including addition of
sulfate groups and inversion of the
carboxyl group on carbon 5 of Dglucuronic acid to yield L-iduronic acid.

Heparin is generated by
hypersulfation of heparan sulfate,
whereas hyaluronan is unsulfated.
The squiggly lines represent
covalent bonds that are oriented
either above (D-glucuronic acid) or
below
(L-iduronic acid) the ring.

 Most GAGs in the ECM are bound to proteins

to form proteoglycans or mucoproteins.
Numerous GAGs (1-200 per molecule,
average length of 800 monosaccharide
units) are attached to a core protein and
different kinds of proteoglycans can be
made by varying the combination of core
proteins and GAGs.
Proteoglycans (MW of~ 1 million) can be
individual or attached to long hyaluronate
molecules to form complexes (as in
cartilage).
They can be embedded in the plasma
membrane or covalently linked to
membrane phospholipids or bound to

 Proteoglycans and collagen may bind to

receptor proteins (often integrins) which are
reinforced by adhesive glycoproteins, such as
fibronectins and laminins, to anchor cells to the
ECM.
GAGs in CT are highly sulfated which attracts
water of hydration. They trap water (up to 50x
their weight) to act as extracellular sponges
resistant to physical forces in cartilage and
joints.
If fluid is injected into CT, it remains localized,
walled off by a viscous ground substance. This
property acts as barrier to the spread of
bacteria that gains access to the tissues.
Some bacteria secrete hyaluronidase
(Staphylo/ Strepto/ Pneumococci), and
collagenase (Clostridium perfringens) that

EDEMA is a condition characterized by

accumulation of excess tissue fluid.

 Edema accompanies pathological conditions that

cause:
Increased hydrostatic pressure
in capillaries by obstructing
venous blood flow
(e.g.
congestive heart failure)
Decreased colloid osmotic pressure in the blood
caused by lack of blood proteins (e.g.
starvation)
Increased hydrostatic pressure in the tissue
caused by blockage of lymphatic
drainage by parasites or tumor cells
Increased colloid osmotic pressure in
the tissue caused by excessive
accumulation of GAGs in
the matrix.

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 Principal producers of collagen fibers are fibroblasts;

epithelial and smooth muscle cells also secrete their
own type-IV collagen.
Most numerous CT matrix, running in all directions in a
wavy course; dull and opaque in appearance.
Fibers bundled together branch and anastomose;
individual fibers do not branch.
With the EM, unit fibrils of collagen show periodic cross
striations every 67 nm of their length.

(a) In tendons, type I

fibrils are all oriented in the
direction of the stress
applied to the tendon.
Proteoglycans and type VI
collagen bind noncovalently
to fibrils, coating the
surface. The microfibrils of
type VI collagen, which
contain globular and triplehelical segments, bind to
type I
fibrils and link them together
into thicker fibers. (b) In
cartilage, type IX collagen
molecules are covalently
bound at regular intervals
along type II fibrils. A
chondroitin sulfate chain,
covalently linked to the 2
type IX chains at the flexible
kink, projects outward from
the fibril, as does the
globular N-terminal region.

Interactions of fibrous
collagens with nonfibrous
fibril-associated collagens.

1.INTRACELLULAR free polysomes reading collagen

mRNA attach to the rER, and protocollagen or
precollagen-chains are deposited in the cisternae. Each
chain has about 250 amino acids; every 3rd amino acid is
glycine.
.The signal peptide is clipped off. Proline and lysine
residues within the chains are then hydroxylated in the
ER to form hydroxyproline and hydroxylysine
(unusual amino acids present in large amounts in
collagen).
.Core sugars (galactose and glucose) attach to the
hydroxylysine residues in the ER.
.Each chain is synthesized with an extra length of
peptides known as registration peptides, which ensure
that the appropriate chains assemble in their correct
position in the resulting triple helical molecule called
procollagen.
.Further glycosylation may occur in the Golgi complex,
where procollagen is packaged for secretion. Golgi

1.EXTRACELLULAR- in the extracellular space,

the enzyme procollagen peptidase cleaves the
registration peptides from procollagen,
converting it to tropocollagen.
.Catalyzed by lysyl oxidase, these become
aligned in staggered fashion to form collagen
fibers, possibly under the control of adjacent
fiber-producing cells.
.The turnover of collagen is slowest in tendons,
fastest in loose CT. Macrophages and
neutrophils break down old collagen, and
replaced by fibroblasts.
.As humans age, extracellular collagen
becomes increasingly cross-linked, & turn-over
slows down in CT.

Because collagen synthesis depends on the

expression of several genes and on several
post-translation events, many human diseases
are associated with faulty collagen synthesis.
Progressive systemic sclerosis- excessive
accumulation of collagen (fibrosis) in almost all
organs
Keloid- local swelling caused by abnormal amounts
of collagen that form in scars of skin
Ehlers-Danlos type IV- aortic/ intestinal rupture
due to faulty transcription of collagen type III
Ehlers-Danlos type VII- increased articular
motility due to decreased procollagen peptidase
activity
Scurvy- ulceration of gums, hemorrhages due to
lack of Vit. C, a cofactor for proline hydroxylase
Osteogenesis imperfecta- spontaneous fractures

YELLOW or ELASTIC FIBERS

Form gentle curves or spirals at their free ends
when released from tension
Do not form bundles; individual fibers branch
and anastomose to form networks
They can be stretched to 150% of their length
without breaking, but lose their resiliency with
advancing age.
Appears yellowish, highly refractile,
homogenous and are not made up of fibrillar
subunits that are visible with the light
microscope.
Each fibril is made up of still smaller fibrils
united by a small amount of ground substance.
These smaller microfibrils have periodic cross
bandings.

Synthesis and Assembly of Elastin:

1.Intracellular- microfibrillar proteins containing
mostly hydrophilic amino acids, and proelastin
(contains large amounts of the hydrophobic amino
acids glycine, proline and valine, thus accounting
for elastins insolubility) are synthesized on rER
and secreted separately.
2.Extracellular- proelastin molecules polymerize
extracellularly to form elastin chains.
Lysyl oxidases then catalyze the conversion of
certain lysine residues of elastin to aldehydes, 3
of which condense with a 4th unaltered lysine
residue to form desmosine and isodesmosine.
These very rare amino acids found in elastin
cross-link individual chains, which then associate
with numerous microfibrils to form a branching
and anastomosing network of elastic fibers.

ARGYROPHYL or RETICULAR
FIBERS
Fibers are not branched,
and are not so wavy as
the collagenous fibers when released from tension.
They are chemically identical to collagen, hence
these fibers are considered as precursors of type I
and III collagen; however, they are thinner and
form delicate networks instead of thick bundles
Chemical characteristics- show affinity to silver
(black) stains, hence argyrophyl; do not yield
gelatin on boiling; not easily dissolved by dilute
acids and alkali; not so easily digested by gastric
juice; not so resistant to solutions of alkaline
pancreatic juice.
Distribution- abundant in regions around blood
vessels, muscle fibers, fat cells, basement
membrane of epithelia, endoneurium, lymphoid
organs and red bone marrow.

 Glycoproteins are
globular proteins to
which shorter, branched
oligosaccharide chains
are covalently bound.
These so-called adhesion
glycoproteins mediate
attachment of cells to
their matrix, influence
the state of
differentiation of cells,
and organization of their
cytoskeleton.
Examples are fibronectin,
laminin, thrombospondin,
chondronectin and
fibrillin.

GLYCOPROTEINS

 FIBRONECTINS, a family of
closely related glycoproteins, are
soluble in body fluids (blood),
insoluble in the ECM and partially
soluble at the cell surface.
The fibronectins bind cells to the
matrix and guide cellular
movement.
The RGD (arginine-glycineaspartate) sequence binds to the
integrin fibronectin receptor.
The fibronectins bind cells to the
ECM by bridging cell-surface
receptors to the ECM.
The intracellular cytoskeleton will
align with the extracellular
fibronectin to detemine cell shape.
In many kinds of cancer, cells
unable to make fibronectins loose
shape and detach from the ECM to
become malignant.

 During cell movement (as

during embryogenesis),
pathways of fibronectins
guide cells to their
destinations.
Soluble plasma
fibronectin promotes
blood clotting by direct
binding of fibrin.
Fibronectins guide
immune cells to wounded
areas and thus promote
wound healing.

 LAMININS bind
cells to the basal
lamina of epithelial
and connective
tissues, and to their
surrounding muscle
cells, fat cells, and
Schwann cells.
The basal lamina
serves as a
structural support
for tissues and as a
permeability barrier
to regulate
movement of both
cell and molecules.

Laminin is a very
large protein
comprised of three
proteins that form a
cross. The domains of
laminin bind type IV
collagen, heparin,
heparin sulfate,
entactin and laminin
receptor proteins in
overlying cells to
allow bridging
between the cells and
the ECM. Progeria
(early onset of aging),
is possibly due to a
defective laminin.

 THROMBOSPONDIN - activated plateletproduct. It binds to fibrinogen, plasmalogen

and its activator; a participant in blood clotting.
Its function is poorly understood.
CHONDRONECTIN- a component of cartilage
matrix that mediates attachment of
chondrocytes to their matrix
FIBRILLIN- a nonsulfated glycoprotein
speculated to be essential for normal
development. It is often associated with elastic
fibers or with epithelial basal laminae.
Marfan syndrome is due to a defective
fibrillin gene on chromosome 15,
characterized by excessively long arms and
legs and progressive dilatation and fatal
rupture of the ascending aorta (Pres.
Abraham Lincoln).

The DGC comprises 3

subcomplexes: dystroglycan,
sarcoglycan/ sarcospan of
integral membrane proteins; and
the cytosolic adapter comprising
dystrophin, other adapter
proteins, and signaling molecules.
Through its O-linked sugars,
dystroglycan binds to components
of the basal lamina, such as
laminin. Dystrophin- the protein
defective in Duchenne muscular
dystrophy, links dystroglycan to the
actin cytoskeleton, and dystrobrevin
links dystrophin to the sarcoglycan/
sarcospan subcomplex. Nitric oxide
synthase (NOS) produces nitric
oxide, a gaseous signaling
molecule, and GRB2 is a component
of signaling pathways activated by
certain cell-surface receptors.
Mutations in dystrophin, other DGC
components, laminin, or enzymes
that add the O-linked sugars to
dystroglycan disrupt the DGCmediated link between the exterior
and the interior of muscle cells and
cause muscular dystrophies.

Molecular Connections Between

ECM and Disease: Muscular
Dystrophy

Schematic model of the

dystrophin glycoprotein complex
(DGC) in skeletal muscle cells.