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THE EXTRACELLULAR
MATRIX

Lecture 14

Essential Cell Biology 3rd Ed Chapter 17

Molecular Biology of the Cell Chapter 19
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LECTURE 10: OBJECTIVES

Identify the major components of the Extracellular Matrix
(ECM) and describe their molecular structure, roles and
distribution in specific tissues
Describe the structure of integrins and their interaction with
specific components of the ECM.
Describe clinically importance of Ehlers-Danlos syndrome

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EXTRACELLULAR STRUCTURES
What are they?
Cell wall; Extracellular Matrix

How are they derived/formed?

Secretions from the cell

What are their components?

fibrous proteins, polysacharides, glycoproteins etc.

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THE EXTRACELLULAR MATRIX (ECM):

ANIMAL CELLS
An insoluble network consisting of secreted macromolecules
(proteins and polysaccharides)
Composed of 3 types of molecules:
STRUCTURAL PROTEINS/FIBERS: collagen and elastins
ADHESIVE PROTEINS: laminins and fibronectins (adhesive
glycoproteins)
MATRIX: proteoglycans and hyaluronic acid

Molecules secreted by cells into their immediate environment

Organized meshwork, dynamic, constantly remodeled
Cell adhesion
Cell migration
Epithelial sheets/tubes
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THE EXTRACELLULAR MATRIX (ECM):

TWO FORMS
Is comprised of 2 major forms
1. Interstitial matrix
2. Basement membrane (Basal
Lamina)

Figure 19-43 Molecular Biology of the Cell ( Garland

Science 2008)

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THE EXTRACELLULAR MATRIX (ECM):

MOLECULAR STRUCTURE AND ROLES
Collagen
Proteoglycan
Aggrecan
+hyaluronan

Fibronectin
Laminin

Figure 19-41 Molecular Biology of the Cell ( Garland Science 2008)

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ECM COMPONENTS: COLLAGEN

Most abundant component of
ECM 25%

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ECM COMPONENTS: COLLAGEN AND

ELASTIN
Structural proteins/fibers
Collagen and Elastin

Skin, bone, cartilage, muscle

tensile strength
What kinds of tissues contain elastins?
-arteries, lungs, intestines
elasticity

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ECM COMPONENTS: GROUND SUBSTANCE

Ground substance consists largely of proteoglycans and
hyaluronic acid.
Proteoglycans are ver y large macromolecules, consisting of a
core protein to which many glycosaminoglycan (GAG) molecules
are covalently attached, somewhat like the bristles around the
stem of a bottle brush.
FUNCTIONS TO:

trap water
resistant to compression return to original shape

occupy space
link to collagen fibers
form network
in bone combined with calcium hydroxyapatite, calcium carbonate

Cell adhesion
embryonic migration
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ECM COMPONENTS:
GAGS
10% by weight but fills most of
space
unbranched polysaccharide chains
disaccharide subunits
amino sugar

Four groups
hyaluronan
chondroitin sulphate, dermatan
sulphate
heparan sulphate, heparin
keratan sulphate

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ECM COMPONENTS: GAGS

Many of the sugars in GAGs have
sulfate and carboxyl groups,
which makes them highly
negatively charged.
The high density of negative
charges attracts water, forming a
hydrated gel.
This gel permits the rapid
diffusion of water-soluble Figure 19-60b
molecules but inhibits the
movement of large molecules and
bacteria.
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Molecular Biology of the Cell ( Garland Science 2008)

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Figure 19-60a Molecular Biology of the

Cell ( Garland Science 2008)
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GAGS AND SIGNALING

This gel permits the rapid diffusion of water-soluble molecules
but inhibits the movement of large molecules and bacteria.
proteoglycans can regulate the activities of secreted signaling
molecules
they can bind growth factors
which can either enhance or inhibit their signaling activity

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ECM COMPONENTS:
ADHESIVE GLYCOPROTEINS - FIBRONECTIN
The principal function of the
fibronectin is to connect cells
to matrices that contain
fibrillar collagen.
At least 20 different forms of
fibronectin have been
identified.
All of them arise from
alternative splicing of a single
fibronectin gene.

The soluble forms of

fibronectin are found in tissue
fluids.
The insoluble forms are
organized into fibers in the
extracellular matrix.
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ECM COMPONENTS:
ADHESIVE GLYCOPROTEINS - FIBRONECTIN
dimer connected at Cterminal S-S linkages
rigid and flexible domains
cell binding segment RGDS
binds receptor in
membrane
domains bind
heparin sulphate
Collagen
hyaluronic acid
fibronectin (selfassociation)

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ECM COMPONENTS:
ADHESIVE GLYCOPROTEINS - FIBRONECTIN
cell adhesion
migration pathways
branching
Differentiation
basal laminae
under skin and between
organs
blood
clotting process, link to
fibrin

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ECM COMPONENTS:
ADHESIVE GLYCOPROTEINS - LAMININ
cross-shaped glycoprotein
3 polypeptides a, b1, b2
carbohydrate (S-S bonds)
Separate binding domains
collagen IV
heparin
heparin sulphate
cell binding
cell specific binding

Figure 19-42a Molecular Biology of the Cell (

Garland Science 2008)

cell surface receptor

Side chain linking forms the basis of
Basal laminae held close beneath
the basal ends of the epithelial cells
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Figure 19-42b Molecular Biology of the Cell (

Garland Science 2008)

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ECM COMPONENTS:
THE BASAL LAMINA FIXED ECM
Basement membrane (Basal Lamina)

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Figure 19-40 Molecular Biology of the Cell ( Garland Science 2008)

FIBROBLAST CELLS
Fibroblasts: common cell
type in connective tissue
Secretes ECM rich in
collagen
Migrates and proliferates
readily in wounded tissue
and in tissue culture

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Fibroblast surrounded by collagen

Fibrils in the skin of embryonic chick

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BASAL LAMINA
A selective barrier of the
movement of cells and a filter
for molecules
Important in regeneration of
tissue following damage/
trauma
Basal lamina usually survives
although cells are damaged/killed
Acts as a scaffold to which
regenerating cells can migrate
maintains original tissue
architecture
Figure 19-44 Molecular Biology of the Cell ( Garland Science
2008)
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ECM COMPONENTS:
CONNECTIVE TISSUE
Connective tissue (CT) forms an extensive compartment in the
body and can be considered as the "glue" that holds the body
together.
Very diverse and ranges in consistency from the fluidity of
blood to the gel-like softness of areolar connective tissue to
the hardness of bone.
Connective tissue consists of cells and extracellular material
secreted by some of those cells.
Unlike the other basic tissues (epithelia, muscle, nervous), the
cells in CT may be widely separated from one another within
the extracellular matrix.
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THE STRUCTURE OF INTEGRINS

Virtually all animal cells express
integrins.
They are the most abundant and widely
expressed class of extracellular matrix
protein receptors.

Some integrins associate with other

transmembrane proteins.
Integrins are composed of two
distinct subunits, known as and
chains.
The extracellular portions of both
chains bind to extracellular matrix
proteins
The cytoplasmic portions bind to
cytoskeletal and signaling proteins.

MOST INTEGRINS ARE ECM PROTEIN

RECEPTORS
In vertebrates, there are
many and integrin
subunits.

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M.B. Srichai and R. Zent

These combine to form at

least 24 different
heterodimeric receptors.

over time or
in response to different
environmental conditions

Active
(extended)

Active
(Clustered)

Extrinsic ligand
Outside-in
(collagen, laminin, fibronectin)

Ligand

D E

Most cells express more

than one type of integrin
receptor.
The types of receptor
expressed by a cell can
change:

Inactive
(bent)

D E

E D

salt bridge
FA complex
kindlin
PIPKIJ

talin

Intrinsic ligand
Insideout
(talin, kindlins)

Talin kindlin
vinculin D-actinin
FAK Cas Src paxillin
ILK

Assembly of the actin cytoskeleton

Activation of signaling pathways

Fig. 2.2 Integrin structure of D subunits lacking the I-domain. a Integrins in their unbound, inactive
resting state. In this state integrins are in a bent conformation and the transmembrane and
cytoplasmic regions are closely associated. b Once activated by talins and kindlins, there is separation of the cytoplasmic and transmembrane subunits and extension of the integrins extracellular
domains. Extracellular ligand binding can occur in this conformation. c When activated integrins
bind to ligand, they cluster at the plasma membrane. Clustering is necessary to send intracellular
signals to form tight focal adhesions (FA), important for actin cytoskeletal assembly and activation
of further downstream signals to control various cellular functions

Transmembrane Domain
The transmembrane (TM) domains of integrins are single spanning structures
comprised of ~2529 amino acid residues that form D-helical coiled coils that
either homo- or heterodimerize (Adair and Yeager 2002). Unlike integrin extracellular
domains, no high-resolution experimental X-ray crystal structures are available for
the TM domain of any integrin heterodimer, and much of the structural data are
based on NMR analysis. Structural information from the DIIbE3 heterodimer TM
domains have only recently been solved in their entirety (Adair and Yeager 2002;

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INTEGRINS ARE SIGNALING RECEPTORS

Integrins are signaling receptors
that control both:
cell binding to extracellular matrix
proteins
intracellular responses following
adhesion

Integrins have no enzymatic

activity of their own.
Instead, they interact with adaptor
proteins that link them to signaling
proteins.

INTEGRINS ARE SIGNALING RECEPTORS

Two processes regulate the
strength of integrin binding to
extracellular matrix proteins:

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affinity modulation

D E

avidity modulation
E

varying the clustering of receptors

They can result from changes:

at the cytoplasmic tails of the
receptor subunits or
in the concentration of
extracellular cations

Active
(extended)

Active
(Clustered)

Extrinsic ligand
Outside-in
(collagen, laminin, fibronectin)

Ligand

varying the binding strength of

individual receptors

Changes in integrin receptor

conformation are central to both
types of modulation.

Inactive
(bent)

D E

E D

salt bridge
FA complex
kindlin
PIPKIJ

talin

Intrinsic ligand
Insideout
(talin, kindlins)

Talin kindlin
vinculin D-actinin
FAK Cas Src paxillin
ILK

Assembly of the actin cytoskeleton

Activation of signaling pathways

Fig. 2.2 Integrin structure of D subunits lacking the I-domain. a Integrins in their unbound, inactive
resting state. In this state integrins are in a bent conformation and the transmembrane and
cytoplasmic regions are closely associated. b Once activated by talins and kindlins, there is separation of the cytoplasmic and transmembrane subunits and extension of the integrins extracellular
domains. Extracellular ligand binding can occur in this conformation. c When activated integrins
bind to ligand, they cluster at the plasma membrane. Clustering is necessary to send intracellular
signals to form tight focal adhesions (FA), important for actin cytoskeletal assembly and activation
of further downstream signals to control various cellular functions

Transmembrane Domain
The transmembrane (TM) domains of integrins are single spanning structures
comprised of ~2529 amino acid residues that form D-helical coiled coils that
either homo- or heterodimerize (Adair and Yeager 2002). Unlike integrin extracellular
domains, no high-resolution experimental X-ray crystal structures are available for
the TM domain of any integrin heterodimer, and much of the structural data are
based on NMR analysis. Structural information from the DIIbE3 heterodimer TM
domains have only recently been solved in their entirety (Adair and Yeager 2002;
Lau et al., 2008a, 2008b, 2009). The DIIb TM domain is a 24 residue D-helix
followed by a backbone reversal that lacks a significant helix tilt (Lau et al. 2008a).

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INSTINCTIVELY !
In inside-out
signaling, changes in
receptor conformation
result from
intracellular signals
that originate
elsewhere in the cell.
For example, at
another receptor

In outside-in
signaling, signals
initiated at a receptor
are propagated to
other parts of the cell.
For example, upon
ligand binding

DISEASES OF THE ECM:

EHLERS-DANLOS SYNDROME
Insufficient collagen
Ehlers-Danlos syndrome
rubber-man
skin and tendons easily stretched
contortionists often suffer from this
disease

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Ehlers-Danlos syndrome is seen in

approximately 1 out of every 5,000
people within the general
population.
It affects the connective tissue of
the joints, blood vessels and skin
providing for loose/flexible joints
(as in double-jointed individuals),
fragile blood vessels (bleeding) and
stretchy skin that bruises easily.

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ADHESION TO THE ECM REQUIRES

INTEGRINS

Figure 19-46 Molecular Biology of the Cell ( Garland Science 2008)

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