Beyond the Cell: Extracellular Structures, Cell Adhesion and Cell Junctions

Extracellular Matrix - an organized network of extracellular materials that is present beyond the immediate vicinity of the plasma membrane -in animal cells, it takes on a remarkable variety of forms in different tissues -has different roles: 1. provide structural support 2. it also influences properties such as tissue excitability, cell shape and movement and development of specialized cellular characteristics

Functions of ECM
Scaffold to stabilize the physical structures of tissues Regulate cell behavior Influence cells survival, development, migration, proliferation, shape and function

Connective tissue underlying an epithelial cell sheet. It consist largely of ECM that is secreted by the fibroblasts.

ECM is always consists of the same three classes of molecules: 1. collagens and elastins- structural proteins which give the ECM its strength and flexibility 2. proteoglycans (protein-polysaccharide complex)-provide the matrix in which the structural molecules are embedded 3. fibronectins and laminins- adhesive glycoproteins - attaches cells to the matrix
Table 11. Extracellular Structures of Eukaryotic Cells
Kind of Organism Extracellular Structure Structural Fiber Components of hydrated matrix Adhesive molecules

Animals
Plants

Extracellular Matrix(ECM)
Cell wall

Collagens and elastins

cellulose

proteoglycans
Hemicelluloses and extensins

Fibronectins and laminins

pectins

Collagens are responsible for the strength of the extracellular complex

Collagens are:
insoluble, extracellular glycoproteins found in all animals the most abundant proteins in the human body They are essential structural components of all connective tissues, such as cartilage bone tendons ligaments fascia skin 19 types of collagens have been found (so far) in humans. The major ones are: Type I. The chief component of tendons, ligaments, and bones. Type II. Represents more than 50% of the protein in cartilage. It is also used to build the notochord of vertebrate embryos.

Type III. Strengthens the walls of hollow structures like arteries, the intestine, and the uterus. Type IV. Forms the basal lamina of epithelia. (The basal lamina is often called the basement membrane, but is not related to lipid bilayer membranes.) A meshwork of Type IV collagens provides the filter for the blood capillaries and the glomeruli of the kidneys. The other 15 types are probably equally important but they are much less abundant

Primary Structure of Collagens The basic unit of collagens is a polypeptide consisting of the repeating sequence (glycine (Gly) - X - Y)n

where X is often proline (Pro) and Y is often hydroxyproline (proline to which an -OH group is added after synthesis of the polypeptide). Secondary and Tertiary Structure The resulting molecule twists into an elongated, left-handed helix (NOT an alpha helix). When synthesized, the N- terminal and Cterminal of the polypeptide have globular domains, which keep the molecule soluble. As they pass through the endoplasmic reticulum (ER) and Golgi apparatus,

The molecules are glycosylated. Hydroxyl (-OH) groups are added to the "Y" amino acid. S-S bonds link three chains covalently. The three molecules twist together to form a triple helix. In some collagens (e.g., Type II), the three molecules are identical (the product of a single gene). In other collagens (e.g., Type I), two polypeptides of one kind (gene product) assemble with a second, quite similar, polypeptide, that is the product of a second gene.
When the triple helix is secreted from the cell (usually by a fibroblast), the globular ends are cleaved off. The resulting linear, insoluble molecules assemble into collagen fibers. They assemble in a staggered pattern that gives rise to the striations seen in this electron micrograph. (Type IV collagens are an exception; they form a meshwork rather than striated fibers.)

Inherited Diseases Caused by Mutant Collagen Genes 1. Brittle-bone disease ("osteogenesis imperfecta") Caused by a mutation in one or the other of the two genes whose products are used to make Type I collagen. Like all the inherited collagen diseases, this one is inherited as a dominant trait. The reason: even though one collagen allele is normal, the assembly of the normal gene product with the mutant product produces defective collagen fibers. 2. Some forms of dwarfism Caused by mutations in a Type II collagen gene. 3. Rubber-man syndrome Caused by a mutations in a Type I collagen gene. The subject has hyperextensible joints, tendons, and skin. (This inherited disorder represents one type of Ehlers-Danlos syndrome.) 4. Another type of Ehlers-Danlos syndrome Is caused by mutations in the gene for Type III collagen. Patients are at risk of rupture of major arteries or the intestine. 2.

Ehlers-Danlos syndrome
Defect in synthesis or structure of fibrillar collagen (mutations have been found in collagen types I, III, IV)
Skin hyperextensibility, joint laxity, fragile skin and vessels, poor wound healing

Elastins. - is a protein in connective tissue that is elastic and allows many tissues in the body to resume their shape after stretching or contracting. Elastin helps skin to return to its original position when it is poked or pinched. - provide flexibility to skin, arteries, and lungs. (These are not glycosylated.)

Stretching a network of elastin molecules

*Collagen and Elastin Fibers are embedded in a matrix of proteoglycans

Proteoglycans - glycoproteins in which a large number of glycosaminoglycans(GAGs) are attached to a single protein molecule. -also called mucoproteins -GAGs (the main carbohydrate components of proteoglycans) *also called mucopolysaccharides *form hydrated gels * have four types: Hyaluronan Chondroitin sulfate and dermatan sulfate Heparan sulfate Keratan sulfate

-contributes to the resilience and pliability of cartilage -important role of proteoglycans is to trap water molecules, thus slowing their flow. Because of their high water content, proteoglycan networks are quite resistant to compression and regain their shape quickly

 Present in all tissues and body fluids Simplest GAG

Hyaluronan is a major polysaccharide component of the ECM

Regular repeating sequence of up to 25,000 disaccharide units No sulfated sugars Not linked to a protein

Abundant in early embryos Resist compressive forces in tissues and joints (lubricating properties) Space filler for embryo development Provide space for cell migration (formation of organs) Wound healing

Functions of Hyaluronan
Resists compressive forces in joints and tissues In osteoarthritis, decreased concentration and decreased molecular weight of intra-articular HA.

FDA approved HA for cosmetic use in humans- 2003

Proteoglycans and Adhesive Glycoproteins Anchor Cells to the Extracellular Matrix

1. 2. Cells are anchored to the ECM by proteoglycan linkages Direct links between the ECM and the plasma membrane are reinforced by a family of ADHESIVE GLYCOPROTEIN -bind proteoglycans and collagen molecules to each other and to receptors on the membrane surface -the two most common kinds: a. Fibronectins b. Laminins *many of the membrane receptors to which glycoproteins bind belong to a transmembrane proteins called integrins.

Fibronectins Bind cells to the Matrix and Guide Cellular Movement Fibronectins
-most common adhesive glycoprotein in the ECM -widely distributed throughout the aimal kingdom -occur in a) soluble form in blood and other body fluids b) as insoluble fibrils in the extracellular matrix c) intermediate form loosely associated with cell surfaces * Though they differ in solubility and location they are encoded by the same gene -Dimer with disulfide bonds at one end *Similar but not identical

*Made from the same gene but differently spliced mRNAs

Structure of a fibronectin dimer

Isoforms of Fibronectin

A. Effects of Fibronectin on Cell Shape

Actin Fibronectin Because the orientation and organization of the cytoskeletal network are important in determining the shape of the cell, fibronectin is thought to be significant in the maintenance of cell shape. (* a possible involvement of fibronectin in cancer is suggested by the observation that many kinds of cancer cells are unable to synthesize fibronectins, with an accompanying loss of normal cell shape and detachment of the cell from the ECM. If such cells are supplied with fibronectin, they often return to their normal shape, recover their ability to bind to the ECM and are no longer malignant).

B. Effects of Fibronectin on Cell Movement

-involved in cellular movement such as cell migrations that occur during early embryonic development. -pathways followed by migrating cells are rich in fibronectin (it suggests that such cells are guided by binding to fibronectin molecules along the way).

C. Effects of Fibronectin on Blood Clotting and Wound Healing

-plasma fibronectin *soluble form of fibronectin *involved in blood clotting and perhaps in wound healing How?: Fibronectin promotes blood clotting because it has several binding domains that recognize fibrin(the blood-clotting protein) and can attach blood platelets to fibrin as the blood clot forms. -also thought to guide cells of the immune system as they

migrate into the wounded areas thus promoting wound healing. Laminins Bind Cells to the Basal Lamina

Laminins - another major adhesive glycoprotein present in the ECM - found mainly in the basal laminae -consist of three different polypeptide chains linked by disulfide bonds and organized into a molecule resembling a cross with three short arms and one long arm - like fibronectin, extracellular laminins can greatly influence a cells potential for migration, growth and differentiation

Basal laminae

Properties of Basal Lamina 1. serves as structural support 2. maintains tissue organization 3. permeability barrier that regulates the movement of molecules as well as of cells eg.* In the kidney- basal lamina functions as filter that allows small molecules but not blood proteins to move from the blood into the urine. * the basal lamina beneath epithelial cells prevents passage of underlying connective tissue cells into the epithelium but permits migration of WBCs needed to fight infections (the effect of basal lamina on cell migration is of special interest because of the recent finding that the surfaces of some cancer cells are enriched in binding sites for laminin- the resulting increase in the ability of CC to bind to BL may facilitate their movement thru the structure and allow them to migrate from one region of the body to another).

-Basal lamina also organizes proteins in the membranes of adjacent cells and is involved in the induction of cellular differentiation and the regeneration of injured tissue

Regeneration experiments indicating the special character of the junctional basal lamina at a neuromuscular junction

Integrins are Cell Surface Receptors that Bind ECM Constituents Integrins - receptor glycoproteins which belong to the family of transmembrane proteins - has a role in integrating the cytoskeleton with the intracellular matrix -very important receptors bec they are the 1o means by which ECM proteins such as collagen, fibronectin, and laminin bind to cells and influence cellular functions as well as structure.

Integrins

The subunit structure of an integrin cell surface matrix receptor

Integrins

Cells regulate the activity of their integrins

Cell Interactions, the Extracellular Matrix, and Cell Wall

Cell-Cell Recognition and Adhesion

-the integrity of multicellular organisms depends on the ability of individual cells to associate in precise patterns to form tissues, organs and organs systems. Such ordered interactions require that individual cells be able to recognize, adhere to and communicate with each other.

Cell-Cell Interactions
Introduction
Cell Adhesion Molecules (CAMs)
Cadherins Selectins N-CAMs

Cell-cell Interaction
Types of Junctions
Tight (occluding) junctions Anchoring junctions
Adherens Desmosome Hemidesmosome Septate junctions (invertebrates only)

Communicating Junctions
Gap Junctions Plasmodesmata (plants only)

INTEGRATING CELLS INTO TISSUES

Multicellular organisms organize their cells to coordinate essential activities. Cells elaborate specialized connections between themselves and with their environment, often clustering to form specialized junctions. Integral membrane proteins, cell- adhesion molecules (CAMs), adhere cells tightly and specifically with cells of the same or similar type and allow for rapid communication. Animal cells also elaborate a complex network of proteins and carbohydrates, the extracellular matrix (ECM), that creates a special environment between the cells.

Schematic overview of major adhesive interactions that bind cells to each other and to the Extracellular Matrix (ECM)

Cell Adhesion Molecules (CAMs)

Binds cells together Homophilic adhesion between cells of a single type Heterophilic binding between cells of different types The cytosol-facing domain if the CAMs are usually connected to elements of the cytoskeleton

Cell-Cell and Cell-Matrix Adhesion Molecules (CAMs) Five major classes: some homophilic; some heterophilic

Transmembrane adhesion proteins link the Cytoskeleton to Extracellular Structures:

Cell-cell adhesions usually mediated by cadherins Cell-matrix adhesions usually mediated by integrins Internal linkage to cytoskeleton is mediated by intracellular anchor proteins

Adhesive Proteins: Cadherins

Calcium dependent cell-cell adhesion glycoproteins Over 40 different forms expressed in tissue specific fashion Antibody to cadherin or removal of calcium leads to dissociation of epithelial cells and failure to form desmosomes and gap junctions Prefers homophilic binding (can thereby mediate cell sorting)

The C-terminal cytoplasmic domain associates with the cytoskeleton N-terminal extracellular domain forms dimers and, through homophilic, interactions forms tetramers Each cadherin has a characteristic tissue distribution

The Cadherin superfamily includes hundreds of different proteins Take their name from their dependence on calcium Extracellular domain containing multiple copies of the cadherin motif Intracellular portions varied Adhesive and signaling functions

Cadherins Mediate Ca2+ - dependent cell-cell adhesion in all animals:

Main adhesion molecules holding cells together in early embryonic tissues

Cadherins
Ca2+ dependent cell-cell adhesion
E-cadherin: epithelial cells N-cadherin: nerve, muscle, fibroblast, & lens cells P- cadherin: placenta, epidermis, breast epithelium VE-cadherin: endothelial cells
Co-receptor for vascular endothelial growth factor (VEGF) which binds to a receptor for tyrosine kinase (signal transduction)

Cadherins mediate homophilic adhesion:

Cadherins of a specific subtype on one cell will bind cadherins of the same type on another cell

Functions of Cadherins
Adherin junction in epithelial cells Connect actin to hold cells together The first cadherin expressed during mammalian development Important in compaction during embryo development Crucial in later stages of vertebrate development Its expression related to the morphogenesis of nerve cells

E-cadherin

N-cadherin

Cell Junctions
-specialized structures of the plasma membrane at the point where two cells come together which are specific means of joining cells in long-term associations to form tissues and organs in multicellular organisms.

INTEGRATING CELLS INTO TISSUES

CELL-CELL ADHESION PROTEINS BIND EXTRACELLULARLY AND ARE ATTACHED TO INTRACELLULAR ATTACHMENT PROTEINS MEMBRANE RECEPTORS BIND INTRACELLULARLY TO CYTOSKELETON

Tissue Organizations
Cells Tissues Organs nerve Blood & blood element Muscle Connective tissue

Functional Significance Adult: Organ Architecture Immune/inflammatory responses Embryonic development Diseases- cardiovascular; cancer; skin

Four Functional Classes of Cell

Junctions in Animal Tissues:
Anchoring junctions
Cell-cell and cell-matrix
Transmit stresses through tethering to cytoskeleton

Occluding junctions
Seal gaps between cells to make an impermeable barrier

Channel-forming junctions (gap junctions)

Link cytoplasms of adjacent cells

Signal-relaying junctions
Synapses in nervous system, immunological

Figure . Summary of the various cell junctions found in animal cell epithelia. This drawing is based on epithelial cells of the small intestine.

Junctions
Occluding junctions: tight junctions Anchoring junctions Cell-cell
Actin Filaments: adhesion belts Intermediate filament: desmosomes

Cell-matrix
focal contacts hemidesmosome

Communicating junctions: gap junctions

Organization of Cell junctions in epithelia

Relative positions of the junctions are the same in all epithelia

Table 7. Junctions Between Animal Cells Type of Junction

Adhesive junctions Desmosome Cell-Cell adhesion Localized points of attachment Localized points of attachment Continuous zones of attachment Localized points of attachment Membranes joined along ridges Connexons (transmembrane proteins with 3nm pores) 25-35 nm Intermediate filaments (tonofilaments) Intermediate Filaments (tonofilaments) Actin microfilaments Actin microfilaments Transmembrane junctional proteins Connexons in one membrane align with those in another to form channels between cells

Function

Features

Intermembrane Space

Associated Structures

Hemidesmosomes

Cell-basal lamina Adhesion Cell-cell adhesion

25-35 nm

Adherens junctions Adhesion belt Focal contact

20-25 nm

Cell-ECM adhesion

20-25 nm

Tight junctions

Sealing spaces between cells

Exchange of ions and molecules between cells

None 2-3 nm

Gap Junction

Anchoring Junctions -occur widely in animal tissue but are esp. prominent In tissues such as heart muscle and skin epithelium that are Subject to mechanical stress and stretching. *despite structural and functional differences among them, all adhesive junctions contain 2 distinct kinds of proteins: 1. intracellular attachment proteins-link the junctions to the appropriate cytoskeletal filaments on the inside of the plasma membrane 2. transmembrane linker proteins-protrude on the outer surface of the membrane and bind cells to each other or to ECM

Cell-Cell Junctions Desmosomes

Buttonlike points of intercellular contact that rivet cells together Desmosome are adhesion plaques attached by cadherinlike transmembrane linker proteins Connect indirectly the intermediate filaments of adjacent cells to form a continuous network throughout the tissue.
Desmosomes attach muscle cells to each other in a muscle. Some "muscle tears" involve the rupture of desmosomes

Hemidesmosomes
Connect the basal surface of epithelial cells to the underlying basal lamina- a specialized mat of ECM at the interface between the epithelium and connective tissue.

Adherens Junctions
-prominent in heart muscle and in the thin layers of tissue that line body cavities and cover body organs 1. Adhesion belt found in epithelial cells which is continuous and form an extensive zone that completely encompass entire cells in a sheet of tissue 2. Focal contact also known as focal adhesion - can attach cells to the ECM

 Occluding junction (Tight Junction) Cell-cell contact Tight junctions in vertebrate Septate junctions in invertebrates Epithelial cells line all the cavities and free surfaces of body The spaces between epithelial cells are tightly sealed separating fluids at either side that have a different chemical compositions A selective permeability barrier specific membrane-bound transport proteins in apical and basolateral surfaces for nutrients

Gap Junctions
- (also called communicating junctions) provide cytoplasmic channels from one cell to an adjacent cell and in this way are similar in their function to the plasmodesmata in plants. Gap junctions consist of membrane proteins that surround a pore through which ions, sugars, amino acids, and other small molecules may pass. Gap junctions are necessary for communication between cells in many types of tissues, including heart muscle, and in animal embryos.

Cell-Cell Junctions Gap Junctions

Connections at the lateral surfaces of cells that allow transport of ions and small molecules (as large as 1-2 nm)

Channels directly link the cytosol of adjacent cells The extent to which channels are open is highly regulated (ex. very high calcium ion conc. closes the channels) In neurons, the passage of ions can lead to propagation of action potentials In smooth muscle, Ca++ transfer can induce contraction Passage of cyclic AMP can lead to signal transduction A hormonal stimulation of one cell can be passed to neighboring cells.

Connexons -at a gap junction, the two plasma membranes from adjacent cells are
joined by tightly packed, hollow cylinders.

6 connexin subunits 2 connexon (one from each cell) form a channel Each connexin crosses the membrane four times Different connexins form junctions that differ in channel size and reguation Hetero-oligomeric connexons can form hetertypic gap junction channels

Plants
cell walls are perforated with channels called plasmodesmata