Physiological Processes where Cell

Adhesion is Important
•  homing of lymphocytes to specific lymphoid organs
•  margination of leukocytes at sites of inflammation
•  platelet deposition at sites of injury
•  metastatic spread of tumor cells
•  embryonic development
•  maintenance of proper tissue growth and
differentiation
•  generation of traction for migration and/or tissue
organization
Most mammalian cells are anchorage dependent
 Most mammalian cells are anchorage dependent

C Chen et al. (Science) 1997

Cell adhesion is the first step; if your cells do not adhere, they will not survive.
 Landmark Tissue Engineering Study

If the cells did not adhere to the scaffold, then they would not have survived.
Good to do first test to see if cells adhere to your scaffold.

From Y. Cao et al. (1997) Plastic and Reconstructive Surgery

Cell Adhesion: The Parts
Cell-Cell adhesion and Cell-Substrate
Adhesion involve SPECIFIC bonds
Major Classes of Adhesion Receptors:
Specific Interactions

•  integrin
–  heterodimers of non-covalently associated α and β
subunits
–  interact simultaneously with ECM and cytoskeleton
–  KD with ECM ~ 10-6 ~10-7 M; affinity can be
modulated
•  immunoglobulin
–  T cell receptor; MHC class I and II; N-CAMs (neural
cell adhesion molecules)
•  cadherin
–  found in intercellular junctional structures
•  selectin
–  play a role in neutrophil binding to endothelial cells
at sites of tissue injury and inflammation
The role of tight junctions in transcellular transport
Tight junctions are key for epithelia to serve as
barriers for solute diffusion
Structure of tight junction between epithelial
cells in small intestine
Current model of tight junction
Anchoring junctions in epitheliurm
Anchoring Junction
A Functional Classification of Cell Junctions

OCCLUDING JUNCTIONS
1. tight junctions (vertebrates only)
2. septate junctions (invertebrates mainly)
ANCHORING JUNCTIONS
Actin filament attachment sites
1. cell-cell junctions (adherens junctions)
2. cell-matrix junctions (focal adhesions)
Intermediate filament attachment sites
1. cell-cell junctions (desmosomes)
2. cell-matrix junctions (hemidesmosomes)
COMMUNICATING JUNCTIONS
1. gap junctions
2. chemical synapses
3. plasmodesmata (plants only)
Figure 19-60. The subunit structure of an integrin cell-surface matrix receptor. Electron micrographs of isolated receptors suggest that the
molecule has approximately the shape shown, with the globular head projecting more than 20 nm from the lipid bilayer. By binding to a matrix
protein outside the cell and to the actin cytoskeleton (via the attachment proteins talin and α-actinin) inside the cell, the protein serves as a
transmembrane linker. The α and β chains are both glycosylated (not shown) and are held together by noncovalent bonds. In the fibronectin
receptor shown, the α chain is made initially as a single 140,000-dalton polypeptide chain, which is then cleaved into one small transmembrane
chain and one large extracellular chain that remain held together by a disulfide bond; this extracellular chain is folded into four divalent-cation-
binding domains. The extracellular part of the β chain contains a repeating cysteine-rich region, where intrachain disulfide bonding occurs; the β
chain has a mass of about 100,000 daltons.
Figure 22-4. Interactions between cell-adhesion molecules during the initial binding and tight binding of T cells, a kind
of leukocyte, to activation endothelial cells. Once a T cell has firmly adhered to the endothelium, it can move (extravasate)
into the underlying tissue. Activation of the endothelium requires signals, such as platelet-activating factor (PAF), that are
released in areas of infection or inflammation; thus extravasation occurs only in such areas. See text for discussion. [Adapted
from R. O. Hynes and A. Lander, 1992, Cell 68:303.]
© 2000 by W. H. Freeman and Company. All rights reserved.
 Cell Surface: Glycocalyx (repellers?)

net negative surface charge

 Cell Adhesion to ECM

COLLAGEN EXTRACELLULAR
MATRIX
•  Direct linkage to collagen or
proteoglycan
–  insertion of fibers into membrane
FIBRONECTIN
–  covalent attachment to membrane
lipid
INTEGRIN
•  Linking glycoproteins
–  fibronectin CELL MEMBRANE

–  laminin

FOCAL
ADHESION

ACTIN

CYTOPLASM
 ECM Function
•  Support for cells
•  Pattern of ECM regulates
–  cell division
–  adhesion
–  motility
•  Development
–  migration
–  differentiation
•  Growth factors
–  Reservoir
•  Dynamic reciprocity (Bissel et al.)
•  Signal transduction
–  ECM molecules
–  cell surface receptors
–  gene expression
 Extracellular Matrix (ECM)
•  Collagens
•  Proteoglycans
•  Glycoproteins
•  Glycosaminoglycans (GAG)
•  Elastin and fibrillin
•  Growth factors and cytokines
 What are these molecules?
•  Collagen -- different types
•  glycoprotein, forms different networks (fibril vs.
network)
•  Proteoglycans -- huge macromolecules
•  Predominantly carbohydrates (glycan) with some
protein
•  Hyaluronic acid backbone
•  Side chains -- glycosaminoglycans , linear
chains or repeating subunits of carbohydrates
–  Specific type for different matrix (cornea vs. ligament vs.
basal lamina)
Extracellular Matrix
 Collagen
•  Tensile
strength and
elasticity
–  tendons,
cartilage, bone
–  half total body
proteins (by
weight)
 Collagen - Protein
•  3 polypeptide (a) chains
– left hand helix, forms fibers
– 14 different (vertebrate) collagens by
different combinations of a-chains
•  numbered I - XIV
•  I, II, III main fibers, flexible
– I
» bone, skin, tendons (90% of all collagen)
– II
» cartilage
 Collagen - Fibers
•  I, II, III have cross striations showing
overlapping packing of individual
collagen molecules
•  IV fine unstriated
–  sheet-like supportive meshwork
–  mature basal laminae
–  tracks for embryonic migration
–  barriers for cell migration
•  V-XII
–  smaller diameter fibers than I-III
–  no striations
 Glycosaminoglycans
•  10% by weight but fill most of space
•  unbranched polysaccharide chains
•  disaccharide subunits
•  amino sugar
•  4 groups
–  hyaluronan
–  chondroitin sulphate, dermatan sulphate
–  heparan sulphate, heparin
–  keratan sulphate
 Glycosaminoglycans
•  Hyaluronan
– or hyaluronic acid, hyaluronate
– Used in development to produce a “cell-
free” space for cells to proliferate and
migrate into
•  heart, cornea
– In adult used in areas of compression
•  tissues, joints
Proteoglycans
Proteoglycans
 Proteoglycans - Function
•  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
 Fibronectin- Structure
•  dimer connected at C-terminal
–  S-S linkages
•  rigid and flexible domains
•  cell binding segment RGDS
–  arg-gly-asp-ser
–  binds receptor in membrane
•  domains bind
–  heparin sulphate, collagen, hyaluronic acid,
gangliosides, fibronectin
Fibronectin Structure
 Fibronectin Function
•  cell adhesion
–  migration pathways
•  blocking fibronectin with Ab
–  prevents neural crest migration
–  extension of axons and dendrites
•  differentiation
•  basal laminae
–  under skin and between organs
•  blood
–  clotting process, link to fibrin
 Laminin - Structure
•  Cross-shaped glycoprotein
– 3 polypeptides a, b1, b2
– carbohydrate (13% by weight)
– separate binding domains
•  collagen IV
•  heparin
•  heparin sulfate
•  cell binding
•  cell specific binding (liver, nerve)
– cell surface receptor
Laminin
 Laminin - Function
•  cell adhesion
–  migration pathways
–  stimulates growth of axons
•  development and regeneration
•  differentiation
•  basal laminae
–  most abundant linking glycoprotein