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:
o structural proteins (collagens and elastins)
o protein-polysaccharide complexes to embed the structural
proteins (proteoglycans)
o 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
D-glucuronic 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 receptor proteins.
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 breakdown matrix components.
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.
Hypothyroidism resulting from this
condition is referred to as myxedema.
C
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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 Interactions of fibrous collagens
direction of the stress applied to the with nonfibrous fibril-associated
tendon. Proteoglycans and type VI
collagen bind noncovalently to
collagens.
fibrils, coating the surface. The
microfibrils of type VI collagen,
which contain globular and triple-
helical 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(IX) chain
at the flexible kink, projects
outward from the fibril, as does the
globular N-terminal region.
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 vesicles release
procollagen into the extracellular space by exocytosis.
[Link]- 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 & cardiac
insufficiency due to mutation in collagen type I
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 elastin’s 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,
GLYCOPROTEINS
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.
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-glycine-aspartate)
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 platelet-product. 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: Molecular Connections Between ECM and
dystroglycan, sarcoglycan/ sarcospan of
integral membrane proteins; and the Disease: Muscular Dystrophy
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 DGC-mediated link between the
exterior and the interior of muscle cells and
cause muscular dystrophies.
Schematic model of the dystrophin
glycoprotein complex (DGC) in skeletal
muscle cells.
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