EXTRACELLULAR MATRIX

Maria Lourdes R. Bernardo, MD, PTRP

Extracellular Matrix (ECM)
• Connective tissue
• a dense network of proteins that lies between cells and
is secreted by the cells within the network

• 3 MAJOR CLASSES OF BIOMOLECULES

– I. Structural Proteins: a. Collagen
b. Elastin
c. Fibrillin-1
– II. Specialized Proteins: a. Fibronectin
b. Laminin
– III. Proteoglycans
Importance of ECM
 forms a substratum upon which cells reside during
normal homeostasis.

 forms “roadways’ within tissues during

morphogenesis and development.

 acts as a barrier to limit cells to specific regions in

tissues

 acts as a depot for growth factors and cytokines

 controls cell behavior

Ia. Collagen
• Most abundant protein in the animal world
• Major component of most connective tissues (25%)
• Provides an extracellular framework with a triple helical
structure
• 28 types w/ 30 polypeptide chains
• COLLAGEN I – major collagens of skin & bone
• COLLAGEN II – major collagen of cartilage

• NONCOLLAGEN COLLAGENS – proteins not classified

as collagens but have collagen-like domains in their
structures
ECM – FIBER TYPES
•COLLAGEN
–TYPE I
–bone, skin, tendon, ligaments, fibrocartilage
–TYPE II
–hyaline cartilage, elastic cartilage
–TYPE III (RETICULAR FIBERS)
–lymphatic tissue
–TYPE IV
–basement membrane
 Structural Classification
• FIBRIL-FORMING COLLAGENS:
– Types I, II, III, V, and XI
• NETWORK-LIKE COLLAGENS:
– Types IV, VIII, and X
• FACIT: Types IX, XII, XIV, XVI, and XIX
• BEADED FILAMENTS: Type VI
• ANCHORING FIBRILS: Type VII
• TRANSMEMBRANE DOMAIN:
– Types XIII and XVII
• OTHERS: Types XV and XVIII
Collagen Type I
• Triple helical structure w/ ~1000 AA
• Each alpha chain is twisted into a left-handed polyproline helix of
3 residues per turn
• 3 alpha chains are then wounded into a right-handed superhelix

• rodlike molecule 1.4nm diameter & 300nm long

• Stabilized by COVALENT CROSS-LINKS formed by LYSYL
OXIDASE
Striking Characteristic: Glycine
residues at every 3rd position
Repeating Structure: (GLY-X-Y)n
(X usually is proline; Y is usually
hydroxyproline)

When proline is in the Y position,

PROLYL HYDROXYLASE
converts it to HYROXYPROLINE.

When lysine is in the Y position,

LYSYL HYDROXYLASE converts
it to HYDROXYLYSINE.
Non-Gly-X-Y Sequences
• Interruptions of the triple helix lacking the
Gly-X-Y repeat sequences resulting in
areas of GLOBULAR STRUCTURE.

– Type IV Collagen – important component of

basement membranes, where it forms a
mesh-like network
Collagen Synthesis
Genetic Diseases
• Ehlers-Danlos Syndrome
• Alport Syndrome
• Epidermolysis Bullosa
• Scurvy
• Menkes Disease
• Osteogenesis Imperfecta
• Chondrodysplasias
Ehlers-Danlos Syndrome
• PRINCIPAL CLINICAL FEATURES:
– hyperextensibility of skin
– abnormal tissue fragility
– increased joint mobility

• 10 types recognized
Ehlers-Danlos Syndrome
• TYPE IV – most serious; tendency for
spontaneous rupture of arteries or the
bowel due to an abnormal type III collagen
• TYPE VI – deficiency of lysyl hydroxylase;
marked joint hypermobility and
a tendency to ocular rupture
• TYPE VIIC – deficiency of procollagen N-
proteinase; formation of abnormal thin,
irregular collagen fibrils, manifested by
marked joint hypermobility and soft skin
Alport Syndrome
• Affects the structure of type IV collagen
fibers
• Abnormalities of the structure of the
basement membrane and lamina densa
• Sign: hematuria
Epidermolysis Bullosa
• Skin breaks and blisters
after a minor trauma

• DYSTROPHIC FORM – mutation in

COL7A1 (affecting type VII collagen)

• EPIDERMOLYSIS BULLOSA SIMPLEX –

mutations in keratin
Scurvy
• Deficiency of ascorbic acid  Impaired
synthesis of collagen due to deficiency of
prolyl and lysyl hydroxylases
• Major signs: bleeding gums, subcutaneous
hemorrhages, poor wound healing
Menke’s Disease
• Deficiency of copper
• Defective cross-linking of collagen and
elastin by the copper-dependent enzyme
lysyl oxidase
• Sign: sparse and coarse hair
Ib. Elastin
• For extensibility and elastic
recoil in tissues
• Synthesized as a soluble
monomer of TROPOELASTIN
• Site: Lungs, large arterial
blood vessels, elastic
ligaments, skin, ear cartilage
• Conditions with deficiency:
Pulmonary emphysema, cutis
laxa, aging of the skin
Williams-Beuren Syndrome
• Deletion in elastin gene (location: 7q11.23)
• Supravalvular aortic stenosis
Collagen vs. Elastin
COLLAGEN ELASTIN
• Many genetic types • One genetic type
• Triple helix • Random coil
• (+) (Gly-X-Y)n repeating • (-) (Gly-X-Y)n repeating
sequence sequence
• (+) hydroxylysine • (-) hydroxylysine
• (+) carbohydrate • (-) carbohydrate
• Intramolecular aldol • Intramolecular desmosine
cross-links cross-links
• (+) extension peptides • (-) extension peptides
Ic. Fibrillin-1
• Large glycoprotein that is a structural
component of microfibrils
• Secreted by fibroblasts and becomes
incorporated into the insoluble microfibril
• It is found in the zonular fibers of the lens,
in the periostium, & associated with the
elastin fibers in the aorta
Marfan Syndrome
• Missense mutation in the gene
(chromosome 15) for fibrillin-1
• Affectation:
– Eyes (ectopia lentis)
– Skeletal system (tall, arachnodactyly,
hyperextensibility of joints)
– CVS (weakness of aortic media  dilation of
ascending aorta)
IIa. Fibronectin
• Major glycoprotein of ECM
• Plays important role in cell adhesion and
cell migration
• Contains Arg-Gly-Asp (RGD) sequence
• 2 TYPES: a. Cell-associated
b. Soluble
 FIBRONECTIN

FIBRONECTIN IS A DIMER, TWO CHAINS LINKED VIA

DISULFIDE BONDING AT THE CARBOXYL TERMINUS.

EACH PROTEIN IN THE DIMER IS 220,000 K IN SIZE,

C
THEREFORE THE INTACT DIMER IS 440,000K IN SIZE

H2N COOH

FN GENE = 50 EXONS, 70KB IN SIZE

TYPE I TYPE II TYPE III VARIABLE

MODULE MODULE MODULE SEQUENCE
Globular Domains
Fibrin
Heparin sulfate
Cell-Surface Receptor
(contains RGDs)
Collagen with separate
domains for types i, ii
and iii)
Hyaluronic acid
 FIBRONECTIN STRUCTURE

RGD IS THE LETTER DESIGNATION FOR THE

AMINO ACIDS ARGININE, GLYCINE AND
ASPARTIC ACID RGD
C

H2N COOH

I1-I5 I9-III1 III8-III10

TYPE I TYPE II TYPE III VARIABLE

MODULE MODULE MODULE SEQUENCE
Basal Lamina
• specialized areas of the ECM that
surround epithelial and some other cells
• PRIMARY COMPONENTS OF THE
BASAL LAMINA:
– 1. 3 PROTEINS: Laminin, entactin, & type IV
collagen
- 2. GAG heparin or heparan sulfate
 IIb. Laminin
• Major protein component of
renal glomerulus and other
basal laminas
• Consists of 3 polypeptide
chains (A, B1 & B2)
(cruciform shape)
• Binding sites for type IV
collagen, heparin, and
integrins on cell surfaces
Pericellular Assembly of
Laminin Matrix

FROM COLOGNATO AND YURCHENKO:DEV.

DYNAMICS 2000
Entactin / Nidogen
• Glycoprotein containing RGD sequence
• Binds to laminin
• Major cell attachment factor
Proteoglycans
• major component of ECM
• glycoproteins that contain covalently
linked GAGs
– CORE PROTEINS – proteins bound
covalently to GAGs
• form the ground substance of the
basement membrane and connective
tissue
 PROTEOGLYCAN VS GLYCOPROTEIN

• PROTEOGLYCAN CARBOHYDRATE LINKAGE

– THE CARBOHYDRATE LINKAGE IS UNIQUE TO PG’S
– (GLUCURONOSYL-B1,3-GALACTOSYL-B 1,3-GALACTOSYL-B 1,4 XYLOSE)

XYL-GAL-GAL-UA LINKER
COMMON TO ALL
PROTEOGLYCANS

SERINE

GLUCURONIC ACID GALACTOSE GALACTOSE XYLOSE

Glycosaminoglycans (GAGs)
• major component of ECM
• Unbranched polysaccharide made up of
repeating disaccharides
• COMPONENTS:
– AMINO SUGAR: D-glucosamine or D-
galactosamine
– URONIC ACID (except in keratan sulfate): L-
glucuronic acid or L-iduronic acid
7 GAGs
• Hyaluronic acid
• Chondroitin sulfate
• Keratan sulfates I & II
• Heparin
• Heparan sulfate
• Dermatan sulfate

• All contains sulfate groups except hyaluronic acid

Hyaluronic Acid/Hyaluronan
Aggrecan • A hyalectin
• first proteoglycan
characterized at the
molecular level.
•Its core protein carries 100+
chondroitin gag chains.
• its amino terminus has a
binding motif enabling it to
bind to hyaluronic acid
(HA-binding).
•link protein stabilizes the
binding of aggrecan to HA.
Hyaluronic Acid
• Unbranched chain of repeating
disaccharide units containing GlcUA and
GlcNAc
• Present in bacteria and in animals and
tissues
• Has a role in permitting tumor cells to
migrate through the ECM
Hyaluronic Acid
a unique gag from either heparan or chondroitin gag’s.
its repeating disaccharide of resembles that of heparan, however
it is never sulfated and is not assembled in the golgi apparatus.
 actively extruded as a polymer by the cell surface,
transmembrane enzyme hyaluronan synthase (ha synthase).
never covalently bound to a core protein

N-ACETYL GLUCURONIC ACID

GLUCOSAMINE
Chondroitin Sulfates
• Prominent components of cartilage and
located at sites of calcification in
endochondral bone and inside neurons
• Provide an endoskeletal structure & help
to maintain their shape
• Repeating disaccharide containing GlcUA
and GalNAc
Keratan Sulfates I & II
• Repeating Gal-GlcNAc disaccharide units
containing sulfate at 6’ position of GlcNAc
or of Gal
Heparin
• Important anticoagulant
• Binds to lipoprotein lipase in capillary walls
• Contains glucosamine (GlcN) & either of
the 2 uronic acids (predominantly IdUA)
• Protein molecule consists of serine &
glycine residues
• Found in granules of mast cells, and in
liver, lungs, and skin
Heparan Sulfate
• Present on cell surfaces
• Contains GlcN
• Predominant uronic acid is GlcUA
 ANY PROTEOGLYCAN CORE PROTEIN
GLYCOSAMINOGLYCAN CHAINS

S S

SERINE

HEPARIN/HEPARAN SULFATE PROTEOGLYCAN

N-ACETYL N-ACETYL GLUCURONIC ACID GALACTOSE XYLOSE

GLUCOSAMINE GALACTOSAMINE
 ANY PROTEOGLYCAN CORE PROTEIN
GLYCOSAMINOGLYCAN CHAINS

CHONDROITIN/DERMATAN SULFATE
XYL-GAL-GAL-UA LINKER
COMMON TO ALL
PROTEOGLYCANS
N
THIS IS A
BRANCH
POINT IN
GLYCOSAMI
SERINE
NO-GLYCAN
ASSEMBLY

N
HEPARIN/HEPARAN SULFATE

N-ACETYL N-ACETYL GLUCURONIC ACID GALACTOSE XYLOSE

GLUCOSAMINE GALACTOSAMINE
 ANY PROTEOGLYCAN CORE PROTEIN
GLYCOSAMINOGLYCAN CHAINS

S S

SERINE

CHONDROITIN/DERMATAN PROTEOGLYCAN

N-ACETYL N-ACETYL GALACTOSE

GLUCURONIC ACID XYLOSE
GLUCOSAMINE GALACTOSAMINE
Dermatan Sulfate
• Widely distributed in animal tissues
• Major GAG synthesized by arterial smooth
muscle cells
• Binds plasma LDL(atherosclerotic lesions)
• Contains IdUA in an α-1,3 linkage to
GalNAc and GlcUA-GalNAc
disaccharides
Some Functions of GAGs &
Proteoglycans
• Structural components of ECM
• Interacts with other proteins
• Polyanions and adds turgor to tissues
• Acts as sieves in the ECM
• Facilitates cell migration
• Acts in compressibility of cartilage in weight-bearing
• Eyes: Corneal transparency, Structural role in sclera
• Anticoagulant
• Components of plasma membranes
• Determine charge-selectiveness of renal glomerulus
• Components of synaptic and other vesicles
• Autoantigens
Biosynthesis of GAGs
• Attachment to core proteins
• Chain elongation
• Chain termination
3 Types of Linkage between
GAGs & their Core Proteins
• An O-glycosidic bond between xylose and
Ser
• An O-glycosidic bond between GalNAc
and Ser
• An N-glycosylamine bond between
G1cNAc and the amide nitrogen of Asn
Mucopolysaccharidoses
• Autosomal recessive inborn errors of
metabolism
• Results from deficiency of enzymes that
degrade GAGs
(lysosomal hydrolases:
endoglycosidases,
exoglycosidases, and
sulfatases)
Major Features of
Mucopolysaccharidoses
• Chronic progressive
• Multisystem affectation
• Organomegaly
• Dysostosis multiplex
• Abnormal facies
• Abnormal hearing, vision, CVS, & mental
development
Laboratory Tests Used in the
Dx of Mucopolysaccharidosis
• Urinalysis
• Assays of suspected enzymes
• Tissue biopsy
• Specific gene tests
• Prenatal diagnosis
Hyaluronidase
• Important enzyme involved in catabolism
of both hyaluronic acid and chondroitin
sulfate
• Widely distributed endoglycosidase that
cleaves hexosaminidic linkages
Bone
• A dynamic structure that
undergoes continuing cycles
of remodeling
• COMPONENTS: proteins (type I collagen)
hydroxyapatite (calcium)
MAJOR CELLS IN BONE REMODELING:
– Osteoclasts (bone resorption)
– Osteoblasts (bone deposition)
Metabolic & Genetic
Disorders in the Bones
• Osteogenesis imperfecta (brittle bones)
• Osteopetrosis (marble bone disease)
• Osteoporosis
Cartilage
• consists of a dense network of collagen fibers and elastic fibers
embedded in chondroitin sulfate.
• HYALINE CARTILAGE – major type
• TYPE II COLLAGEN – principal protein
• AGGRECAN – major proteoglycan
• 3 DOMAINS OF CORE PROTEINS:
– DOMAIN A – hyaluronic acid
– DOMAIN B – keratan sulfate chains
– DOMAIN C – chondroitin sulfate chains
• CHONDRONECTIN – attachment of type II collagen to
chondrocytes
3 Types of Cartilage.
• HYALINE CARTILAGE
– most abundant, but weakest
– fine collagen fibers embedded in a gel-type matrix
– affords flexibility and support and reduces
friction and absorbs shock in the joints
• FIBROCARTILAGE
– bundles of collagen fibers in its matrix
– strongest of the three types of cartilage
• ELASTIC CARTILAGE
– contains a threadlike network of elastic fibers
– provides strength and elasticity and maintains
the shape of certain organs
Chondrodysplasias
• Mixed group of
hereditary disorders
affecting cartilage
• Manifested by short-
limbed dwarfism &
numerous skeletal
deformities
• Mutations in COL2A1
gene
Achondroplasia
• most common cause of short-limbed dwarfism
• short limbs, normal trunk size, macrocephaly,
and a variety of other skeletal abnormalities
• molecular basis: mutation of nucleotide 1138 in the gene
encoding fibroblast growth factor receptor 3 (FGFR3) on
chromosome 4  replacement in FGFR3 of glycine
(codon 380) by Arginine (rendering it inactive) 
defective function of FGFR3  abnormal development
and growth of cartilage leading to short-limbed dwarfism
and other features
Thank you!!