Extracellular Matrix & Cell Junctions

Collagen
Elastic fibers
Fibronectins & integrin
Laminins
Basal lamina
Proteoglycans
Hemidesmosomes & focal adhesions
Occluding & Adherens junctions
Desmosomes
Gap junctions

Cell-cell interactions
Physical interactions between cells are necessary in a
multicellular organism for:
- tissue cohesion and strength
- coordinating behavior of cells in a tissue
- facilitating changes in cell behavior

Interactions include:
- physical attachment to extracellular structures,
especially extracellular protein fibers (and other
elements of extracellular matrix)
- cell-cell contact via cell junctions of several types
- cell membrane receptors for intercellular signal
molecules
 Fibers of extracellular matrix (ECM): collagens
Collagens = major class of extracellular matrix proteins
Form fibrous structures of different types:
- high-strength fibers of various thicknesses (many organs)
- flexible thinner beaded fibers – attach to ECM
components
- fibrous networks to which layers of cells attach
- membrane-
spanning fibers
that are parts
of some cell-
matrix junctions

Collagen fiber structure

Secreted proteins produced in many cell types: 29 different
sequences, most common is Type I
- primary sequence highly repetitive (most of molecule)
- forms a (non-a) helix that intertwines with two other
collagen molecules: coiled coil
=> collagen _________

Triple helices align with overlap

to make a collagen ______
- small gaps => striations in EM
- covalent cross-links stabilize
Fibrils align and assemble into
collagen fibers of various size
 Collagen synthesis & assembly – part 1
Secreted protein: synthesis begins on rER as
___________ polypeptide with extra N & C sequences
(green in figure)
- polypeptide moves into lumen of rER
- covalent modifications: hydroxylation & glycosylation
=> glycoprotein
- formation of coiled-coil, but
not including N & C-terminal
sequences
- disulfide bonds form
between polypeptides in
triple helix (outside helix)
- packed into large tubular

Collagen synthesis & assembly – part 2

Moves to Golgi for modifications of sugars
Movement of modified coiled-coil from Golgi to plasma
membrane in a fibripositor vesicle
- specific proteases cut off
N & C sequences to generate
____________
- tropocollagens self-associate
to start forming fibrils
- secretion by fusion of large
vesicle, complete association
to fibrils and fibers
- covalent crosslinking of amino
acid sidechains
 Elastic fibers
Elastic fibers, unlike collagen, can stretch and snap back to
their unstressed length
- major protein is elastin, formed from crosslinked
________ polypeptide
- covered by a protein
(microfiber) sheath
Tropoelastin contains
multiple _________
domains that fold into
compact domains
=> very insoluble
Tension unfolds these
domains, stretching
out the fiber

Collagen-cell interactions: fibronectin

Flexible multi-domain ECM glycoprotein fibronectin binds to
collagen fibers and other proteins
- disulfide-linked dimer: two binding sites
- contains binding sites for: fibronectin =>oligomerization
and _________ = plasma membrane protein – links
collagen to cells
Oligomer-
forming
 Integrins
Transmembrane heterodimeric proteins in two conformations:
- inactive: do not bind collagen or other ECM components
or other cell surface proteins
- activated by internal or external signal => reach out
to bind fibronectin or other ECM proteins

Integrin – fibronectin networks

Fibronectins oligomerize by N-terminal domains, making a
network on cell surface – bind many ECM components
- integrins bind (via adapter proteins) to actin fibers in
cytoplasm – moved by ____________ interactions
 Laminins – versatile class of bridging proteins
Highly varied class of ECM glycoproteins that interact with
numerous other proteins (ECM and cell-surface)
______________ of over 10 different polypeptides
- central coiled-coil couples, provides length
- globular domains
have varied binding
sites for other
proteins: cross-links
other ECM fibers
Roles in ECM networks,
must be cleaved by
proteases to allow cell
migration

Basal lamina/basement membrane

Sheet-like ECM network in contact with many cell types
- connects basal layer of cells to underlying tissue
Laminin and collagen networks
Roles: structural, selective filter, binding other components
 Laminins in basal lamina
One specific type of laminin (111) forms a network, linked to
integrins in plasma membrane and type IV collagen
network in ECM

Laminin also binds to

a ___________
(perlecan) =
highly glycosylated
polypeptide

Proteoglycan coupled to
collagen network

Proteoglycans
Very highly ____________ polypeptides (mostly
polysaccharide), free in ECM (or bound to cell surface)
Sugars: acidic or sulfated, formed into long chains =
glycosaminoglycans (GAGs), attached to a polypeptide
core
High _______________:
repels itself: spreads out &
attracts & holds water
=> dense hydrated gel
Also bind to other proteins
=> adhesion, often by GAG
heparan sulfate
 Hyaluronan (HA)
Very large GAG – no polypeptide
- binds noncovalently to proteoglycans - huge complexes
Major function: resistance to ____________
- major component of
cartilage
Also binds cell surface
receptors, increases
space between cells

Hemidesmosomes
Epithelial cell-ECM junction at basal lamina
- _________ couple a protein complex (plaque) in
cytoplasm to laminins in basal lamina, which are coupled
to collagen network
- cytoplasmic plaque coupled to _______ filaments (IFs)
 Hemidesmosomes & focal adhesions
Integrins also form basis of focal contacts / focal
complexes / focal adhesions
- attach to fibrous ECM elements, clustering integrins –
reversible for migrating cells
- in cytosol, various proteins bind integrins to ______
filaments: force transduction

Junctional complex in epithelia

Usually ordered from apical to basal surfaces:
Occluding/tight
Adherens
Desmosomes

Communicating/
gap junctions

Hemidesmosomes on
basal surface only
 Tight/occluding junctions
Tight junctions – number of intersecting bands on lateral
surface of epithelia : zonlula occludens (ZO)
- __________________between the cells, limit
movement of molecules (&
viruses) between cells
- 3 TM proteins identified:
claudins, occludins and
junctional adhesion molecules
- proteins bind actin in cytosol

Paracellular transport
Transport between compartments via the
extracellular space between cells
- mainly blocked (and regulated) by
_________
- allow select ions pass rapidly
- individually break open, allowing other
molecules to pass (slowly)
Also divide cell membrane into
apical and basolateral
domains, limiting movement
of lipids and proteins
between the two
 Adherens junctions
In epithelia, form a band (zonula adherens, ZA) just basal
to the ZO
- linked by transmembrane proteins: ________ that
bind neighboring cell cadherins: homophilic binding

Cytosolic domain of cadherins

binds proteins (catenins in
epithelial ZA) that strongly
bind actin filaments
=> resists stress (at apical
region)
=> allows cell shape change
by action of myosin

Remodeling of cells in development

Early development: transformation of cell layers into 3D
structures
Cells must change shape and position in response to signals
Adherens junctions allow myosin to pull on actin filaments
at cell edge
to reshape
entire layer
of cells
Cell communication
coordinates
changes
 Adherens junctions in other cells
Most prominent in epithelial ZA, but also used in other cells
that require adhesion – more limited in size

Desmosomes
Cell adhesion complexes in various cells, most prominent in
epithelia and between cardiac & smooth muscle cells
“Spot welds” on lateral surface that consist of:
- transmembrane proteins (desmo-names) of the ______
family
- cytoplasmic anchoring
proteins forming a
dense plaque (double
layer at hi-res,
plako-names) that bind
- _______________
to resist mechanical
stress on cell
 Gap junctions: direct cell-cell communcation
Small channels connect cells in some tissues to allow ions
and small molecules to equilibrate between cells :
____________
Connexons = paired hexamers of connexin monomers that
form a tight seal in the intercellular space
- can be open or closed, depending on cell conditions
- one protein (homo-hexamer) or mixed (hetero-hexamer)

NCAMs: neuronal cell adhesion molecules

Nerve cells express adhesion proteins (NCAMs) that bind
_________ and also proteoglycans
- modulate strength of binding by modification by neg-
charged sugars (poly-sialic
acids, PSA)
 Selectins: attracting circulating cells
White blood cells (leukocytes) are drawn out of blood to
infection sites by binding their surface __________ to
selectins on the surface of endothelial cells

Lectins = proteins that bind

specific oligosaccharides

Selectins expressed as an
inflammatory response
- bind oligosaccharide
(sialyl Lewis) on
leukocutes, slow and stop