AL-FARABI KAZAKH NATIONAL UNIVERSITY

Higher School of Medicine

Intercellular interactions. Glycocalyx. Cell adhesion, cell contacts.

Extracellular matrix
 Learning outcomes:
• identify and describe the main mediators of intercellular contacts at the physical level.
• compare connective and epithelial tissues depending on their cellular and ECM components
• describe the role of the glycocalyx in the formation of intercellular influences
• describe the general scheme of cell contacts
• name the main types of cell contacts
• define adhesive contacts, describe the role of cadherins in adhesive junctions and desmosomes
• explain the role of cadherins in cell distribution
• describe which components of the cytoskeleton are associated with cadherins
• define tight junctions, describe the functional and structural properties of tight junctions, explain what a
junctional complex is
• define a gap junction, describe the functional and structural properties of gap junctions
• explain what a connexon is
• describe the structural and functional properties of the extracellular matrix;
• explain the molecular basis of the organization of cells in tissues;
• explain the relationship between the cell-cell/cell-extracellular matrix interaction and the cell cycle.
• explain the role of cell-cell and cell-ECM intercellular contacts in resistance to mechanical stress
• tell what types of cells secrete ECM; list the main components of the ECM
• describe the chemical properties of glycosaminoglycans; give examples of GAGs
• explain the chemical structure of proteoglycans; give examples of proteoglycans
• explain what collagen is; give an overview of the classification of collagen
• define a glycoprotein
• describe the structural and functional properties of the basal lamina
• list the main components of the basal lamina and their structure; laminins
 Literature
• Cooper M. Geofrey The Cell: A Molecular Approach // 8th edition - Oxfod Press,
2018-819p.
• Bruce Alberts, Dennis Bray Karen Hopkin and et all. –Essential Cell Biology, 4th
Edition [text]: textbook. hard cover. Garland Science, 2013 - 865 p
Most of the cells in multicellular organisms are organized into cooperative assemblies called
tissues, to perform a common function, such as the nervous, muscle, epithelial, and connective
tissues found in vertebrates.

Different tissues can be organized into an organ, again to perform one or more specific functions.
The coordinated functioning of many types of cells and tissues permits the organism to move,
metabolize, reproduce, and carry out other essential activities
• The assembly of distinct tissues and their organization into organs are determined by molecular interactions
at the cellular level . These interactions would not be possible without the temporally, spatially, and
functionally regulated expression of a wide array of adhesion molecules.
• Cells in tissues can adhere directly to one another (cell-cell adhesion) through specialized membrane
proteins called cell-adhesion molecules (CAMs), which often cluster into specialized cell junctions.
• Cells in animal tissues also adhere indirectly (cell-matrix adhesion) through the binding of adhesion
receptors in the plasma membrane to components of the surrounding extracellular matrix (ECM), a complex
interdigitating meshwork of proteins and polysaccharides secreted by cells into the spaces between them.
Some adhesion receptors can also function as CAMs, mediating direct interaction between cells.
• both types of adhesions are intrinsically associated with the cytoskeleton and cellular signaling pathways.
As a result, a cell’s surroundings influence its shape and functional properties (“outside-in” effects);
likewise, cellular shape and function influence a cell’s surroundings (“inside-out” effects). Thus
connectivity and communication are intimately related properties of cells in tissues.
 Cell-cell interactions
• Direct interactions between cells, are critical to the development and function of multicellular
organisms. Some cell–cell interactions are transient, such as the interactions between cells of
the immune system and the interactions that direct white blood cells to sites of tissue
inflammation. In other cases, stable cell–cell junctions play a key role in the organization of
cells in tissues.
Anchoring junctions and tight junctions perform the key task of holding the tissue
together. gap junctions, permits the rapid diffusion of small, water-soluble molecules
between the cytoplasms of adjacent cells. Along with anchoring and tight junctions, gap
junctions help a cell communicate with its environment.
 Tight junctions
• These specialized junctions form a barrier that seals off body cavities such as the
intestinal lumen and separates the blood from the cerebral spinal fluid of the central
nervous system (i.e., the blood-brain barrier). Tight junctions prevent the diffusion of
macromolecules and, to varying degrees, small water-soluble molecules and ions across
an epithelium via the spaces between cells. They also help establish and maintain the
polarity of epithelial cells by preventing the diffusion of membrane proteins and
glycolipids between the apical and the basolateral regions of the plasma membrane,
ensuring that these regions contain different membrane components
• Tight junctions are composed of thin bands of plasma membrane proteins
that completely encircle the cell and are in contact with similar thin bands
on adjacent cells
• The occludin and claudin are principal integral membrane proteins found in tight junctions,
which are arranged in strands along the lines of the junction to create the seal
• A group of junction adhesion molecules (JAMs) have also been found to contribute to
homophilic adhesion and other functions of tight junctions.
• The extracellular domains of rows of occludin, claudin, and JAMs in the plasma membrane of
one cell apparently form extremely tight links with similar rows of the same proteins in an
adjacent cell, creating a tight seal.
• Two additional transmembrane proteins are incorporated into the tight junctions: tricellulin,
which has four membranespanning helices, as do occludin and claudins, and angulins,which
have a single transmembrane helix and one extracellular immunoglobulin domain and which
appear to be required for the assembly of tricellulin where the cells intersect
 Cytosolic adapter proteins and their connections to the
cytoskeleton are critical components of tight junctions.
• Several adapter proteins are associated with tight junctions, including the zonula
occludens (ZO) proteins ZO-1, ZO-2, and ZO-3, which not only interact with
occludin, claudin, and other adapter and signaling proteins but also mediate
association with actin fibers. These interactions appear to stabilize the linkage
between occludin and claudin molecules that is essential for maintaining the
integrity of tight junctions. ZO proteins can also function as adapters for adherens
junctions and gap junctions
 Gap Junctions
• The activities of individual cells in multicellular organisms need to be closely
coordinated. This can be accomplished by signaling molecules that are released
from one cell and act on another. However, within an individual tissue, such as the
liver, cells are often linked by gap junctions, which provide direct connections
between the cytoplasms of adjacent cells. Gap junctions are regulated channels
through the plasma membrane that, when open, allow ions and small molecules
(less than ~1000 daltons) to diffuse between neighboring cells.
• Most cells in animal tissues— including epithelial cells, endothelial cells, and the
cells of cardiac and smooth muscle—communicate by gap junctions
• Vertebrate gap junctions are composed of connexins, a family of structurally related
transmembrane proteins with molecular weights between 26,000 and 60,000. Each
vertebrate hexagonal particle consists of twelve noncovalently associated connexin
molecules: six form a cylindrical hemichannel, called a connexon, in one plasma
membrane that is joined to a connexon in the adjacent cell membrane, forming a
continuous aqueous channel (diameter ∼14 Å) between the cells.

• Each individual connexin molecule has four membrane-spanning α helices with a

topology similar to that of claudin, resulting in 24 transmembrane α helices in each
connexon hemichannel.
• Some cells express a single connexin that forms homotypic connexons. Most cells,
however, express at least two connexins; these different proteins can assemble into
heteromeric connexons, which in turn form heterotypic gap-junction channels. Diversity
in channel composition leads to differences in channel permeability. The permeability of
gap junctions is regulated by posttranslational modification of connexins (e.g.,
phosphorylation) and is sensitive to changes in environmental conditions such as
intracellular pH and Ca2+ concentration, membrane potential, and the intercellular
potential between adjacent interconnected cells (“voltage gating”). The N-termini of
connexins appear to be especially important in the gating mechanism
 Anchored junctions
• Adherens junctions, desmosomes, and focal adhesions are found in many
different types of cells; hemidesmosomes appear to be restricted to
epithelial cells
 • Cell–cell adhesion is a selective process such that cells adhere only to other
cells of specific types. Such selective cell–cell adhesion is mediated by
transmembrane proteins called cell adhesion molecules, which can be
divided into four major groups:

Cell adhesion mediated by the selectins, integrins, and most cadherins requires Ca2+, Mg2+,
or Mn2+, so many adhesive interactions between cells are divalent cation-dependent.
• The selectins mediate transient interactions between leukocytes and
endothelial cells or blood platelets. There are three members of the select in
family: L-selectin, which is expressed on leukocytes; E-selectin, which is
expressed on endothelial cells; and P-selectin, which is expressed on
platelets. The selectins recognize cell surface carbohydrates
Leukocytes leave the circulation at sites of tissue inflammation by interacting with the endothelial
cells of capillary walls. The first step in this interaction is the binding of leukocyte selectins to
carbohydrates (oligosaccharide ligands) on the endothelial cell surface. The oligosaccharide
contains N-acetylglucosamine (GlcNAc), fucose (Fuc), galactose (Gal), and sialic acid
(N-acetylneuraminic acid, NANA). This step is followed by more stable interactions between
leukocyte integrins and intercellular adhesion molecules (ICAMs)—members of the Ig
superfamily—on endothelial cells.
The primary CAMs in adherens junctions and desmosomes belong to the cadherin
family. Many different types of cells in the widely diverse tissues of these animals
use cadherins to mediate adhesion and communication, the detailed requirements
for which may differ for different types of cells and tissues. Members of the
cadherin superfamily can also control cell morphology, such as the assembly and
tight packing of microvilli on the apical surfaces of some epithelial cells
There are said to be over 100 different types of cadherins found in vertebrates, which
can be classified into four groups:

✔ classical
✔ desmosomal
✔ protocadherins
✔ unconventional
The classical cadherins include E-, N-, and P-cadherins, named for the type of
tissue in which they were initially identified (epithelial, neural, and
placental, respectively)

Each classical cadherin molecule contains a single transmembrane domain, a

relatively short C-terminal cytosolic domain, and five extracellular
“cadherin” domains (called EC1–EC5)
 At a typical cell–cell junction, an organized array of cadherin molecules functions like
Velcro to hold cells together. Cadherins on the same cell are thought to be coupled by
sideto-side interactions between their N-terminal head regions, resulting in a linear
array like the alternating green and light green cadherins on the lower cell shown here.
These arrays are thought to interact with similar linear arrays on an adjacent cell (blue
cadherin molecules, top cell). The linear arrays on one cell are perpendicular to those
on the other cell, as indicated by the red arrows. Multiple perpendicular arrays on both
cells interact to form a tight-knit mat of cadherin proteins
• The adhesiveness of cadherins depends on the presence of extracellular Ca2+; it is
this property (calcium adhering) that gave rise to their name

• The extracellular domains are necessary for Ca2+ binding and cadherin-mediated
cell-cell adhesion. Classical cadherin–mediated adhesion entails both cis lateral
clustering (intracellular) and trans adhesive (intercellular) molecular interactions.
The binding of three Ca2+ at each of the sites located between the cadherin repeats
stabilizes the elongated and curved structure of the extracellular domain.
 • The C-terminal cytosolic domain of classical cadherins is linked to the actin
cytoskeleton by adapter proteins. These linkages are essential for strong adhesion,
as a moderate increase in tension generated by the actin cytoskeleton induces the
formation of larger clusters of cadherins and stronger intercellular adhesion

Some of the increased cadherin-mediated adhesion that accompanies increased force applied by the actin
cytoskeleton appears to be mediated by one of the adapter proteins, α-catenin, a mechanosensor that links
cadherin to actin filaments and changes shape (stretches out) when subjected to force. β-catenin—another
common adapter protein that links classical cadherins to actin filaments — dramatically reduces
cadherin-mediated cell-cell adhesion
• Desmosomes contain two specialized cadherins, desmoglein and desmocollin,
whose cytosolic domains are distinct from those in the classical cadherins.
• The cytosolic domains of desmosomal cadherins bind to adapter proteins such as
plakoglobin (similar in structure to β-catenin) and plakophilins, and these bind to a
member of the plakin family of adapters, called desmoplakin.
• These adapters form the thick cytoplasmic plaques that are characteristic of
desmosomes. The desmoplakins directly mediate plaque binding to intermediate
filaments.
• To be stably anchored to solid tissues and organs, simple columnar epithelial sheets
must be firmly attached via their basal surfaces to the underlying ECM (basal
lamina). This attachment occurs via adhesion receptors called integrins, which are
located both within and outside of anchoring junctions called hemidesmosomes.
Hemidesmosomes comprise integral membrane proteins linked via cytoplasmic
adapter proteins (e.g., plakins) to keratin-based intermediate filaments.
Integrins function as adhesion receptors and CAMs in a wide variety of epithelial and
nonepithelial cells, mediating many cell-matrix and cell-cell interactions. . In vertebrates,
at least 24 integrin heterodimers, composed of18 types of α subunits and 8 types of β
subunits in various αβ heterodimeric combinations, are known. A single type of β chain
can interact with any one of several different types of α chains, forming distinct integrins
that bind different ligands.
• Parts of both the α subunit and the β subunit of an integrin molecule contribute to
the primary extracellular ligand binding site . Like that of other adhesion molecules,
the cytosolic region of integrins interacts with adapter proteins, which in turn bind
to the cytoskeleton and to intracellular signaling molecules. Most integrins are
linked via adapters to the actin cytoskeleton, including two of the integrins that
connect the basal surface of epithelial cells to the basal lamina via the ECM
molecule laminin. Some integrins, however, interact with intermediate filaments.
 Extra cellular matrix
• Although animal cells are not surrounded by cell walls, most animal cells
in tissues are embedded in an extracellular matrix that fills the spaces
between cells and binds cells and tissues together
• There are several types of extracellular matrices, which consist of a variety
of secreted proteins and polysaccharides
• Many functions of the ECM and, indeed, some features of the assembly of
the ECM require transmembrane adhesion receptors, including the integrins,
that bind directly to ECM components and that also interact, through adapter
proteins, with the cytoskeleton.

• Molecules abundant in the ECM of all tissues are:

Proteoglycans

Collagen fibers

Soluble multi-adhesive proteins

(laminin, fibronectin)
 Basal lamina
• One type of extracellular matrix is exemplified by the thin, sheetlike basal
laminae upon which layers of epithelial cells rest.
• In animals, most organized groups of cells in epithelial and nonepithelial
tissues are underlain or surrounded by the basal lamina, a sheet-like
meshwork of ECM components usually no more than 60–120 nm thick.
The basal lamina is structured differently in different tissues.
Tissue regeneration

Embryonic developement
Form a tight barrier that limits diffusion of
molecules between the blood and the brain and
kidney
Blood filter

Protect the cell membranes from damage during

muscle contraction
• Most of the ECM components in the basal lamina are synthesized by the
cells that rest on it. Four ubiquitous protein components, each of which
comprises multiple, distinct, repeating domains, are found in basal laminae

basal lamina is linked to cells by adhesion receptors, including integrins in

hemidesmosomes, which bind to laminin in the basal lamina. The other side of the
basal lamina is anchored to the adjacent connective tissue by a layer of collagen fibers
embedded in a proteoglycan-rich matrix
• Laminin, the principal multi-adhesive matrix protein in basal laminae, is a heterotrimeric
protein comprising α, β, and γ chains. At least 16 laminin isoforms in vertebrates are
assembled from 5 α, 3 β, and 3 γ chains. Each laminin isoform exhibits a distinctive pattern of
tissue- and developmental stage–specific expression. Many laminins are large, cross-shaped
proteins (molecular weight of about 820,000), although some are Y or rod shaped.

Globular domains at the N-terminus of each

subunit bind to one another and thus mediate
the self-assembly of laminins into mesh-like
networks.
Five globular LG domains at the C-terminus
of the laminin α subunit mediate
Ca2+-dependent binding to cell-surface
laminin receptors, including certain integrins
Some of these interactions are via negatively
charged carbohydrates on the receptors. LG
domains are found in a wide variety of other
proteins and can mediate binding to steroids
and proteins as well as carbohydrates.
Laminin is the principal basal laminal ligand
of integrins.
• Type IV collagen is, together with laminin, a principal structural component of all basal laminae
and can bind to adhesion receptors, including some integrins. Collagen IV is one of at least 28
types of collagen in humans that participate in the formation of distinct ECMs in various tissues.
• There are also at least 20 additional collagen-like proteins !!
• All collagens are trimeric proteins made from three polypeptides
• All or parts of the three-stranded collagen molecule can twist together into a special triple helix
called a collagenous triple helix.
• Within a helical segment, each of the three α chains twists into a left-handed helix, and the three
chains then wrap around one another to form a right-handed triple helix
• The unique properties of each collagen
isoform are due mainly to differences
in
• (1) the number and lengths of the
collagenous triple-helical segments;
• (2) the segments that flank or interrupt
the triple-helical segments and that
fold into other kinds of
three-dimensional structures;
• (3) covalent modification of the α
chains (e.g., hydroxylation,
glycosylation, oxidation,
cross-linking).
 • Perlecan, the major secreted proteoglycan in basal laminae, consists of a large
multidomain core protein (∼470 kDa) to which polysaccharides are covalently
attached. The core protein is made up of multiple repeats of five distinct domains,
including laminin-like LG domains (3 copies), EGF-like domains (12 copies), and
Ig domains (22 copies). The many globular repeats give it the appearance of an
approximately 200-nm-long string of pearls when visualized by electron
microscopy; hence the name perlecan

Perlecan contains three types of covalent polysaccharide chains: N-linked chains, O-linked
chains, and glycosaminoglycans (GAGs)
 Connective tissue
• Connective tissue, such as tendon and cartilage, differs from other solid
tissues in that most of its volume is made up of extracellular matrix rather
than cells. This ECM is packed with insoluble protein fibers.
 ECM in connective tissue has several key components, some of
which are found in other types of tissues as well:
✔ Collagens, trimeric molecules that are often bundled together into fibers (fibrillar
collagens)
✔ Glycosaminoglycans (GAGs), specialized linear polysaccharide chains of specific
repeating disaccharides that can be highly hydrated and confer diverse binding and
physical properties (e.g., resistance to compression)
✔ Proteoglycans, glycoproteins containing one or more covalently bound GAG chains
✔ Multi-adhesive proteins, large multidomain proteins often comprising many copies
(“repeats”) of a few distinctive domains that bind to and cross-link a variety of adhesion
receptors and ECM components
✔ Elastin, a protein that forms the amorphous core of elastic fibers

Collagen is the most abundant fibrous protein in connective tissue. Rubber-like elastin
fibers, which can be stretched and relaxed, are also present in deformable sites (e.g.,
skin, tendons, heart). The fibronectins, a family of multi-adhesive matrix proteins, form
their own distinct fibrils in the ECM of most connective tissues
• About 80–90 percent of the collagen in
the body consists of fibrillar collagens
(types I, II, and III), located primarily in
connective tissues
• Its fundamental structural unit is a long
(300-nm), thin (1.5-nm-diameter) triple
helix consisting of two α1(I) chains and
one α2(I) chain, each 1050 amino acids
in length. The triple stranded molecules
pack tightly together and wrap around
one another, forming microfibrils that
associate into higher-order polymers
called collagen fibrils, which in turn
often aggregate into larger bundles called
collagen fibers
• Fibrillar collagens are secreted proteins,
produced primarily by fibroblasts in the
ECM
• Collagens differ in the structures of the fibers they form and in how these fibers are
organized into networks. Of the predominant types of collagen found in connective
tissues, type I collagen forms long fibers, whereas networks of type II collagen are
more mesh-like

Interactions of fibrillar collagens with fibril associated collagens. (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 type I
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 α2(IX) chain at the flexible kink, projects outward from the fibril,
as does the globular N-terminal region.
• In tendons, for instance, the long type I collagen fibers connect muscles to bones and must
withstand enormous forces. Because type I collagen fibers have great tensile strength, tendons
usually can be stretched without being broken. Indeed, gram for gram, type I collagen. is
stronger than steel. Two quantitatively minor fibrillar collagens, type V and type XI,
co-assemble into fibers with type I collagen, thereby regulating the structures and properties of
the fibers. Incorporation of type V collagen, for example, results in smaller-diameter fibers.
• Type I collagen fibrils are also used as the reinforcing rods in the construction of
bone. Bones and teeth are hard and strong because they contain large amounts of
dahllite, a crystalline calcium- and phosphate-containing mineral. Most bones are
about 70 percent mineral and 30 percent protein, the vast majority of which is type I
collagen. Bones form when certain cells (chondrocytes and osteoblasts) secrete
collagen fibrils that are then mineralized by deposition of small dahllite crystals.
• The fibrils of type II collagen, the major collagen in cartilage, are smaller in diameter than
type I fibrils and are oriented randomly in a viscous proteoglycan matrix. The rigid collagen
fibrils impart strength to the matrix and allow it to resist large deformations. Type II fibrils are
cross-linked to matrix proteoglycans by type IX collagen, another fibril-associated collagen.
The interrupted triple-helical structure of type IX and related collagens prevents them from
assembling into fibrils, although they can associate with fibrils formed from other collagen
types and form covalent cross-links to them.
• proteoglycans play an important role in cell-ECM adhesion. Proteoglycans
are a subset of secreted or cell-surface glycoproteins containing covalently
linked, specialized polysaccharide chains called glycosaminoglycans
(GAGs).
• GAGs are long linear polymers of specific repeating disaccharides. GAG
consist of
• uronic acid (d-glucuronic acid or l-iduronic acid) or d-galactose +
N-acetylglucosamine or N-acetylgalactosamine
• With the exception of hyaluronan, these sugars are
modified by the addition of sulfate groups.
Consequently, GAGs are highly negatively charged.
• In general, GAGs are strongly hydrophilic and tend to
adopt highly extended conformations, which occupy a
huge volume relative to their mass. Thus GAGs act as
effective “space fillers” in the extracellular matrix of
connective tissues.
• Even at very low concentrations, GAGs form
hydrophilic gels: their multiple negative charges attract
a cloud of cations, such as Na+, that are osmotically
active, causing large amounts of water to be sucked
into the matrix. This gives rise to a swelling pressure,
which is balanced by tension in the collagen fibers
interwoven with the proteoglycans
 • GAGs are usually covalently linked to core proteins to form proteoglycans, which
are extremely diverse in size, shape, and chemistry. Typically, many GAG chains
are attached to a single core protein, which may in turn be linked at one end to
another GAG, creating an enormous aggregate resembling a bottlebrush, with a
molecular weight in the millions. Proteoglycans can contain as few as one or as
many as more than 100 GAG chains attached to serine residues of a core protein

Proteoglycans perform many sophisticated functions in addition to providing hydrated space around
cells. They can form gels of varying pore size and charge density that act as filters to regulate the
passage of molecules through the extracellular medium. They can bind secreted growth factors and
other proteins that serve as extracellular signals for cells. They can block, encourage, or guide cell
migration through the matrix.
• Thanks for attention!