CHAPTER 30
Cellular Adhesion
A ll cells interact with molecules in their environment,
in many cases relying on cell-surface adhesion proteins
to bind these molecules. Multicellular organisms are
particularly dependent on adhesion of cells to each other
and the extracellular matrix (ECM). During development,
carefully regulated genetic programs specify cell-cell
and cell-matrix interactions that determine the architec-
ture of each tissue and organ. Some adhesive interactions
are stable. Muscle cells must adhere firmly to each
other and to the connective tissue of tendons to transmit
force to the skeleton (see Chapter 39). Skin cells must
also bind tightly to each other and the underlying con-
nective tissue to resist abrasion (see Fig. 35.6). On the
other hand, many cellular interactions are transient
and delicate. At sites of inflammation, leukocytes roll
along endothelial cells lining small blood vessels and
then interact weakly with the ECM as they migrate
through connective tissue (look ahead to Figs. 30.13
and 30.14).
Cells use a relatively small repertoire of adhesion
mechanisms to interact with matrix molecules and
each other. A conceptual breakthrough came when
comparisons of amino acid sequences showed that
most adhesion proteins fall into five large families
(Fig. 30.1). Within each of these distinctive families,
ancestral genes duplicated and diverged during evolu-
tion, giving rise to adhesion proteins with the many
different specificities required for embryonic develop-
ment, maintenance of organ structure, and migrations
of cells of our defense systems. General principles
emerged from characterizing a few examples. Several
important adhesion proteins fall outside the five major
families, and additional families may emerge from con-
tinued research.
Many adhesion proteins were named before they
were classified into families. Tables 30.1 through 30.5
are designed to help the reader with the challenging
nomenclature. Many adhesion proteins are named
Immunoglobulin
family (ICAM-1) CD54
E-cadherin
Integrin L2
(LFA-1)
CD11a/CD18
P-selectin
CD62P
Mucin
CD43
FIGURE 30.1 CLASSES OF ADHESION PROTEINS. Immuno-
globulin cell adhesion molecules (IgCAMs) have one or more extracel-
lular domains folded like an immunoglobulin domain (orange).
Cadherins have five or more extracellular CAD (cadherin) domains
(maroon) that typically bind the same class of cadherin on a neighbor-
ing cell. Ca2+ stabilizes the interaction of neighboring CAD domains.
Their cytoplasmic tails bind -catenin (blue) and other adapter pro-
teins. Integrins are heterodimers of and subunits that bind a wide
range of matrix molecules by combinations of 1 of 16 -chains and
1 of 8 -chains. Selectins have a Ca2+-dependent lectin (carbohydrate-
binding) domain (green), an epidermal growth factor (EGF)-like domain
(blue), and a variable number of complement regulatory domains
(red). Mucins use multiple carbohydrates for interactions with other
cells. CD numbers refer to the names of representative adhesion
molecules using the clusters of differentiation nomenclature (see the
text). Single transmembrane helices anchor all these receptors to the
plasma membrane. Adapter proteins link the cytoplasmic tails of most
adhesion proteins to the actin cytoskeleton or, in the case of special-
ized cadherins and integrins, to intermediate filaments. ICAM, inter-
cellular adhesion molecule; LFA, lymphocyte functionassociated
antigen. (Modified from van der Merwe PA, Barclay AN. Transient
intercellular adhesion: the importance of weak protein-protein interac-
tions. Trends Cell Biol. 1994;19:354358.)
525
526 SECTION VIII n Cellular Adhesion and the Extracellular Matrix
CD followed by a number. This stands for clusters of
differentiation, a term used to classify cell-surface anti-
gens recognized by monoclonal antibodies. These names
were given without knowledge of the structure or func-
tion of the antigen. Hence, members of the four major
families of adhesion proteins have CD numbers.
This chapter first highlights some general features
of adhesion proteins and then introduces the five
major families: immunoglobulincell adhesion molecules
(IgCAMs), cadherins, integrins, selectins, and
mucins. While learning about each family, the reader
should bear in mind that these receptors rarely act
alone. Rather, they usually function as parts of multi-
component systems. Two examples at the end of the
chapter illustrate the cooperation that is required for
leukocytes to respond to inflammation and for platelets
to repair damaged blood vessels. Chapter 31 on intercel-
lular junctions, Chapter 32 on specialized connective
tissues, and Chapter 38 on cellular motility provide
more examples of cellular adhesion.
General Principles of Cellular Adhesion
First Principle of Adhesion
Cells define their capacity for adhesion by selectively
expressing plasma membrane receptors (cell adhesion
molecules [CAMs]) with selective ligand-binding activ-
ity. Generally, expression of the proper mix of recep-
tors is part of a genetic program for cell differentiation.
In some cases, extracellular stimuli control expression
of adhesion receptors. For example, inflammatory
hormones or endotoxin stimulate endothelial cells to
produce E-selectin on demand.
Second Principle of Adhesion
Many adhesion proteins bind one main ligand, and
many ligands bind a single type of receptor (refer to
Tables 30.1 through 30.5). If this one-to-one pairing
were the rule, adhesion would be simple indeed.
However, many exceptions exist, particularly in the
integrin family of receptors (Table 30.3), which gener-
ally bind more than one ligand, and some ligands,
such as fibronectin, bind more than one integrin. One
can generalize about the ligands for the several families
of CAMs:
Most cadherins prefer to bind themselves, so they
promote the adhesion of like cells. These homophilic
interactions (association of like receptors on two
cells) require Ca2+.
Most IgCAMs bind other cell-surface adhesion pro-
teins. These heterophilic interactions (association
of unlike receptors) may occur between the same or
different cell types.
Selectins bind anionic polysaccharides like those on
mucins. Generally, such interactions bind together
two different types of cells.
A
a
b
Dissociated cells Cluster of cells sorted
of two types with b cells inside
High level
of cadherin
Low level Cluster with cells expressing
of cadherin a high level of cadherin
B on the inside
FIGURE 30.2 SORTING OF EMBRYONIC CELLS. A, When cells
from different tissues are dissociated and mixed together, they spon-
taneously sort themselves into two layers, with the more adherent cells
inside the less-adherent cells. B, Cells with high concentrations of
cadherin sort inside less-adherent cells. (Based on the work of M.
Steinberg, Princeton University, Princeton, NJ.)
Integrins stand apart because they bind a variety of
ligands: matrix macromolecules, such as fibronectin
(see Fig. 29.14) and laminin (see Fig. 39.9); soluble
proteins, such as fibrinogen in blood; and adhesion
proteins on other cells, including IgCAMs and one
cadherin.
Third Principle of Adhesion
Cells modulate adhesion by controlling the surface
density, state of aggregation, and state of activation of
their adhesion receptors. Surface density reflects not
only the level of synthesis but also the partitioning of
adhesion molecules between the plasma membrane and
intracellular storage compartments. For example, endo-
thelial cells express P-selectin constitutively but store it
internally in membranes of cytoplasmic vesicles. When
inflammatory cytokines activate endothelial cells, these
vesicles fuse with the plasma membrane, exposing
P-selectin on the cell surface, where it binds white blood
cells (Fig. 30.13). The importance of surface density is
illustrated by an experiment with mixtures of cells that
express different levels of the same cadherin. Over time,
they sort from each other, with the more adherent cells
forming a cluster surrounded by the less-adherent cells
(Fig. 30.2). Such differential expression of cadherin
determines the position of the oocyte in Drosophila egg
follicles. A variety of extracellular stimuli activate intra-
cellular signaling pathways in lymphocytes, platelets,
and other cells. These pathways enhance or inhibit the
ligand-binding activity of integrins already located on the
cell surface.
Fourth Principle of Adhesion
The rates of ligand binding and dissociation are impor-
tant determinants of cellular adhesion. Many cell-surface

adhesion proteins (including IgCAMs, cadherins, integ-
rins, and selectins) bind their ligands weakly compared
with other specific macromolecular interactions, such
as antigens and antibodies, hormones and receptors, or
transcription factors and DNA. The measured dissocia-
tion equilibrium constants for adhesion receptors are in
the range of 1 to 100M, reflecting high rate constants
(>1s1) for dissociation of ligand. This actually makes
good biological sense. Rapidly reversible interactions
allow white blood cells to roll along the endothelium of
blood vessels (Fig. 30.13). Transient adhesion also allows
fibroblasts to migrate through connective tissue. In
contrast, the interactions of cells in epithelia and muscle
appear to be more stable, perhaps owing to multiple
weak interactions between clustered adhesion pro-
teins cooperating to stabilize adherens junctions and
desmosomes (see Fig. 31.8). The combined strength
of these bonds is said to increase the avidity of the
interaction.
Fifth Principle of Adhesion
Many adhesion receptors interact with the cytoskeleton
inside the cell. Adapter proteins link cadherins and
integrins to actin filaments or intermediate filaments.
These interactions provide mechanical continuity from
cell to cell in muscles and epithelia, allowing them to
transmit forces and resist mechanical disruption.
Sixth Principle of Adhesion
Association of ligands with adhesion receptors activates
intracellular signal transduction pathways, leading to
changes in gene expression, cellular differentiation,
secretion, motility, receptor activation, and cell division.
Signaling through adhesion receptors allows cells to
adjust their behavior based on physical interactions with
the surrounding matrix or cells.
Identification and Characterization
of Adhesion Receptors
The ability of mixed populations of cells to sort into
homogeneous aggregates revealed that mechanisms
exist to bind like cells together. Similar experiments
showed that cells also bind matrix macromolecules, such
as fibronectin, laminin, collagen, and proteoglycans.
Biochemical isolation of the responsible adhesion pro-
teins was challenging, but progressed rapidly once
monoclonal antibodies (see Fig. 28.10) that inhibit adhe-
sion were available. These antibodies provided assays for
purification of adhesion proteins and cloning of their
complementary DNAs (cDNAs). With representatives
from each family in hand, cloning cDNAs for related
proteins was straightforward. Once the first crystal
structures were determined, the structures of other
family members could be modeled using the structures
of shared functional domains.
CHAPTER 30 n Cellular Adhesion 527
Insights about the functions of adhesion receptors
came in several steps. Localization of a protein on spe-
cific cells frequently provided the first clues. Typically,
the expression of each protein is restricted to a subset
of cells or to a specific time during embryonic develop-
ment or both. Next, investigators used specific antibod-
ies to test for the participation of the adhesion protein
in cellular interactions in vitro or in tissues. Both human
genetic diseases and mutations in mice and other organ-
isms produce defects caused by the absence of adhesion
proteins. Blistering skin diseases called pemphigus illus-
trate the serious consequences when pathological auto-
antibodies disrupt adhesion between skin cells expressing
the antigen (see Fig. 31.8). In leukocyte adhesion defi-
ciency, white blood cells lack the 2-integrin that is
required to bind the endothelial cells that line blood
vessels. These defective white blood cells fail to bind to
blood vessel walls or to migrate into connective tissue
at sites of infection. Patients with a bleeding disorder
called Bernard-Soulier syndrome lack one of the adhe-
sion receptors for von Willebrand factor, a protein that
promotes platelet aggregation. Loss of cadherins contrib-
utes to the spread of some cancer cells.
Immunoglobulin Family of Cell
Adhesion Molecules
The IgCAM family includes hundreds of adhesion proteins
that bind ligands on the surfaces of other cells. Some
interactions are homophilic binding to the same IgCAM
on another cell; others are heterophilic with different
IgCAMs, integrins, other proteins or proteins with sialic
acid. These interactions help specify interactions between
different cell types in developing and mature animals.
IgCAMs have one to seven extracellular immunoglobu-
lin domains anchored to the plasma membrane by a
single transmembrane helix (Fig. 30.3 and Table 30.1).
The compact immunoglobulin (Ig) domains consist of 90
to 115 residues folded into seven to nine -strands in two
sheets, usually stabilized by an intramolecular disulfide
bond. The N- and C-termini are at opposite ends of the
Ig domains, so they can form linear arrays. Some nervous
system IgCAMs have three or four fibronectin III (FN-III)
domains (see Fig. 13.13) between the Ig domains and
the membrane anchor. Most IgCAMs consist of a single
polypeptide, but others are multimeric, with two (CD8)
or four subunits (see Fig. 27.8 for the T-cell receptor).
The C-terminal cytoplasmic tails of these receptors
vary in sequence and binding sites for intracellular
ligands. The cytoplasmic domains of the lymphocyte
accessory receptors CD4 and CD8 bind protein tyrosine
kinases required for cellular activation (see Fig. 27.8 for
CD4). The cytoplasmic domains of neuronal IgCAMs
bind PDZ domain proteins or the membrane skeleton
(see Fig. 13.11). An adapter protein links IgCAMs in the
nectin family to cytoplasmic actin filaments.
528 SECTION VIII n Cellular Adhesion and the Extracellular Matrix

A
Differentiated metazoan cells express IgCAMs selec-
1
2 1 1 tively, especially during embryonic development, when
3 2
4 3 2 they contribute to the specificity of cellular interactions
3 2 1 5 5 1 required to form the organs. Neurons and glial cells
4 1 6 6 2
2 1 express specific IgCAMs that guide the growth of
5 3 2 7 7 3 3
neurites, mediate synapse formation, and promote
the formation of myelin sheaths. In adults, interaction of
endothelial cell intercellular adhesion molecule (ICAM)-1
ICAM-1 Nectin ICAM-2 VCAM-1 MAdCAM-1
CD54 CD102 CD106 with a white blood cell integrin is essential for adhesion
and movement of the leukocytes into the connective
B tissue at sites of inflammation (Fig. 30.13).
Like other cell adhesion proteins, IgCAMs participate
in signaling processes. Best understood are interactions
of lymphocytes with antigen-presenting cells during
immune responses. IgCAMs reinforce the interaction of
T-cell receptors with major histocompatibility complex
molecules carrying appropriate antigens on other cells
CD4D1D2 CD4D3D4 CD8/ (see Fig. 27.8). Although individual interactions are
FIGURE 30.3 STRUCTURES OF REPRESENTATIVE IMMUNO- weak, the combination of specific (T-cell receptor) and
GLOBULIN CELL ADHESION MOLECULES. A, Domain maps of nonspecific (CD2 and CD4) interactions with the target
examples with their common names and CD numbers. B, Ribbon dia- cell suffices to initiate signaling.
grams of the lymphocyte coreceptors CD4 (domains 1 and 2 on the left
and domains 3 and 4 in the middle) and CD8. ICAM, intercellular adhe-
sion molecule; MAdCAM, mucosal addressin cell adhesion molecule; Cadherin Family of Adhesion Receptors
VCAM, vascular cell adhesion molecule. (For reference, see Protein Data
Bank [PDB; www.rcsb.org] files 3CD4, 1CID, and 1CD8.) The complex architecture of organs in vertebrates relies
on Ca2+-dependent associations between the cells medi-
ated by more than 80 cadherins (Table 30.2). Their name
derives from calcium-dependent adhesion protein.

TABLE 30.1 Immunoglobulin Family of Cell Adhesion Molecules*

Extracellular Intracellular
Examples Structure Ligands Ligands Expression Functions
CD2 2Ig-1TM LFA-3 (CD58) T cells T-cell activation
CD4 4Ig-1TM Class II MHC Lck T cells, macrophages T-cell coreceptor
CD8 Dimer: 1Ig-1TM Class I MHC Lck Cytotoxic; other T cells T-cell coreceptor
C-CAM 4Ig-1TM Self Liver, intestine, WBCs Cell adhesion
F11 (contactin) 6Ig-4FN-II-1TM Neurons Neurite fasciculation
ICAM-1 5Ig-1TM LFA-1, MAC-1 Epithelia, WBCs WBC adhesion
ICAM-2 2Ig-1TM Endothelium, WBCs
L1 (Ng-CAM) [mouse] 6Ig-3FN-III-1TM Self Ankyrin Neurons, Schwann cells Adhesion
LFA-3 (CD58) 2Ig-1TM or GPI CD2 WBCs, epithelia, Adhesion
anchor fibroblasts
MAG 5Ig-1TM Neurons Glial cells Myelin formation
NCAM 5Ig-3FN-III-1TM Self Neurons, other cells Adhesion
Neurofascin [chick] 6Ig-4FN-III-1TM ? Self Ankyrin Neurites Bundling neurites
PECAM-1 (CD31) 6Ig-1TM Self Platelets, endothelium, Adhesion
myeloid cells
TAG-1 6Ig-4FN-III-GPI anchor ? Self Neurons Neuron migration
VCAM-1 7Ig-1TM WBC 4 integrin Endothelium WBC/endothelium
(regulated) adhesion
CAM, cell adhesion molecule; CD, cellular differentiation antigen; FN-III, fibronectin-III domain; GPI, glycosylphosphatidylinositol; ICAM, intercellular
adhesion molecule; Ig, immunoglobulin domain; Lck, nonreceptor tyrosine kinase; LFA, lymphocyte functionassociated antigen; MAG, myelin-associated
glycoprotein; MHC, major histocompatibility complex; NCAM, neural cell adhesion molecule; PECAM, platelet endothelial cell adhesion molecule; TAG,
transient axonal glycoprotein; TM, transmembrane domain; VCAM, vascular cell adhesion molecule; WBC, white blood cells.
*Hundreds are known.

Partial atomic structure.

 CHAPTER 30 n Cellular Adhesion 529

TABLE 30.2 Cadherin Family of Adhesion Molecules*

Type (Examples) Extracellular Ligands Intracellular Ligands Expression Functions
Classic Cadherins
E-cadherin Self Catenins (actin) Epithelia, others Adherens junctions
N-cadherin Self Catenins (actin) Neurons, muscle, Adhesion
endothelium
R-cadherin Self Catenins (actin) Retina, neurons Adhesion
Desmosomal Cadherins
Desmocollins Self, desmogleins Plakoglobin (desmoplakin, Epithelia Desmosomes
plackophilin, IF)
Desmogleins Self, desmocollins Plakoglobin (desmoplakin, Epithelia, heart Desmosomes
plackophilin, IF)
Atypical Cadherins
T-cadherin Self None (GPI anchor) Early embryos, neurons Intercellular adhesion
Protocadherins
-, -, and Self Fyn tyrosine kinase (some) Vertebrate neurons, Self-avoidance
- Protocadherins other cells
Signaling Cadherins
RET protooncogene Self None Endocrine glands, Intercellular adhesion
neurons

GPI, glycosylphosphatidylinositol; IF, intermediate filament.

*More than 80 are known. Plakoglobin is also known as -catenin.

Genes for cadherin domains appeared in unicellular

precursors of sponges, representing an early step toward A. Desmosome B. Adherens junction
the evolution of metazoan organisms.
Cadherins generally interact with like cadherins on
the surfaces of other cells in a calcium-dependent
fashion, but some cadherins form heterophilic interac-
Cadherins
tions. Homophilic interactions of cadherins link epithelial
Cadherins
and muscle cells to their neighbors, especially at special-
ized adhesive junctions called adherens junctions and
desmosomes (Fig. 30.4; also see Fig. 31.8).
The structural hallmark of the cadherin family is the
CAD domain (Figs. 30.5 and 30.6), which consists of
approximately 110 residues folded into a sandwich
of seven -strands. This fold resembles Ig and FN-III
domains, but appears to be a case of convergent evolu-
tion. N- and C-termini are on opposite ends of CAD
FIGURE 30.4 ELECTRON MICROGRAPHS OF ROD-LIKE
domains. Ca2+ bound to three sites between adjacent CADHERINS CONNECTING THE PLASMA MEMBRANES OF
CAD domains links them together into rigid rods. Without ADJACENT CELLS. Intestinal epithelial cells were prepared by
Ca2+, the domains rotate freely around their linker rapid freezing, freeze-fracture, deep etching, and rotary shadowing.
peptides. A, Desmosome with associated intermediate filaments in the cyto-
Classic cadherins interact head to head through their plasm. B, Adherens junction with associated actin filaments. (Courtesy
N. Hirokawa, University of Tokyo, Japan. Modified from Hirokawa N,
N-terminal CAD1 domains (Fig. 30.6D) forming strong Heuser J. Quick-freeze, deep-etch visualization of the cytoskeleton
trans-interactions with a partner on another cell. beneath surface differentiations of intestinal epithelial cells. J Cell Biol.
Cadherins on the same cell can interact laterally in 1981;91:399409, copyright The Rockefeller University Press.)
cis-interactions. Three-dimensional reconstructions of
electron micrographs of desmosomes show both trans- has a glycosylphosphatidylinositol (GPI) anchor (see
and cis-interactions (Fig. 30.6B). Cadherins are synthe- Fig. 13.10). Cytoplasmic domains vary in size, sequence,
sized with a small domain before the N-terminal and binding sites for associated proteins. The protoon-
interaction strand, which must be removed by proteoly- cogene RET is a cadherin with a cytoplasmic tyrosine
sis to allow binding to another cadherin. kinase domain.
A single -helix links classic cadherins and desmo- Adapter proteins link the cytoplasmic domains of
somal cadherins to the plasma membrane, but T-cadherin cadherins to actin filaments or intermediate filaments
530 SECTION VIII n Cellular Adhesion and the Extracellular Matrix
E-cadherin
ICS
-catenin binding
Desmocollin-1a
ICS
Desmoglein-1
ICS
T-cadherin
Glycosylphosphatidyl-
inositol anchor
Protocadherins
Src-family
kinase binding
RET
Tyrosine kinase
FIGURE 30.5 DOMAIN MAPS OF CADHERINS. All have extra-
cellular CAD domains. A single transmembrane segment anchors five
examples. A glycosylphosphatidylinositol (GPI) tail anchors T-cadherin.
Intracellular cadherin segment (ICS) domains interact with catenin
adapters to link E-cadherin to actin filaments and desmocollin and
desmoglein to intermediate filaments.
to reinforce adhesion and maintain the physical integrity
of tissues (see Fig. 31.8). The cytoplasmic tails of classic
cadherins bind along the entire length of the adapter
protein -catenin (catenin is link in Greek), a long,
twisted coil of 36 short -helices (see Fig. 13.10F).
Monomers of -catenin link -catenin to actin fila-
ments, an interaction strengthened by tension. -Catenin
dimers also stabilize actin filaments. The more compli-
cated cytoplasmic domains of desmosomal cadherins
(desmocollins and desmogleins) interact with -catenin
(a relative of -catenin also called plakoglobin) and
desmoplakin. Desmoplakin links these cadherins to
keratin intermediate filaments (see Fig. 31.8B). The tails
of some cadherins interact with formins, proteins that
nucleate and elongate actin filaments (see Fig. 33.14).
Signaling by Cadherins and Catenins
In addition to helping with the mechanical sorting of
embryonic cells, cadherins produce signals that influ-
ence cellular proliferation, migration, and differentiation.
For example, interactions between cadherins on epithe-
lial cells result in contact inhibition of both growth
and motility (see Figs. 41.3 and 41.11). Rho-family gua-
nosine triphosphatases (GTPases) (see Fig. 33.19)
mediate contact inhibition of movements. The GTPase
Rho stimulates contraction at the contact site, while the
GTPase Rac drives protrusion of the side of the cell
facing away from the contact site. Both cadherins and
Eph receptor tyrosine kinases participate in this reaction
to cell-cell contact. Engagement of cadherins at sites of
contact also stops cellular proliferation through the
Hippo signaling pathway of protein kinases. This pathway
inhibits expression of genes required for cell cycle pro-
gression such as cyclin-dependent kinases (see Fig. 41.3).
The mechanism involves phosphorylation of a transcrip-
tion factor, excluding it from the nucleus.
Contact inhibition of growth and motility suppresses
the spread of cancer cells, so mutations disabling
E-cadherin can contribute to the transition from benign
to invasive malignant tumors. For example, genetic
defects in E-cadherin predispose people to stomach
cancer. The oncogenic tyrosine kinase Src (see Box 27.5)
phosphorylates both E-cadherin and -catenin. This
phosphorylation is associated with loss of adhesion of
epithelial cells, suggesting one way in which transforma-
tion might alter cellular adhesion.
In addition to linking cadherins to actin filaments,
-catenin is an active component in the Wnt signal
transduction pathway that regulates gene expression
during differentiation of embryonic cells (Fig. 30.7).
Extracellular signaling proteins called Wnts regulate the
concentration of -catenin available to regulate gene
expression. The pathway was discovered in Drosophila,
where it helps to determine the polarity of segments in
early embryos. Wnts were named from the original
Drosophila gene Wingless and the mouse protoonco-
gene Int-1. The human genome encodes 29 Wnts.
Most -catenin in cells is bound to cadherins, but a
second pool exchanges between the cytoplasm and the
nucleus, where it recruits transcription factors to regu-
late the expression of genes for cellular proliferation and
tissue differentiation. Cells synthesize -catenin continu-
ously, but it turns over rapidly in resting cells, so little
accumulates in the nucleus. A cytoplasmic complex
controls degradation of -catenin. The complex consists
of two kinasesglycogen synthase kinase (GSK) and
casein kinase-1and the product of the APC gene
(defective in patients with familial adenomatous polypo-
sis coli, giving rise to multiple precancerous polyps in
the large intestine). Phosphorylated -catenin is ubiqui-
tylated and degraded by proteasomes.
Wnts suppress the degradation of -catenin by binding
to seven-helix receptors and another class of receptors
in the plasma membrane. Several steps downstream in
an incompletely characterized pathway, the Wnt signal
inhibits GSK and casein kinase-1. Inhibition of the
kinases stops proteolysis of -catenin, so ongoing synthe-
sis of -catenin raises the concentration that is free to
accumulate in the nucleus. Stem cell proliferation is one
of many developmental events influenced by Wnt signal-
ing and adhesion by cadherins (see Box 41.2). Wnt
binding its receptors also activates a parallel noncanoni-
cal pathway involving Rho-family GTPases that control
the cytoskeleton during cell migration.
 CHAPTER 30 n Cellular Adhesion 531

A. EM data D. Crystal structures

B. 3D reconstruction from EM
E-CAD1

E-CAD2

E-CAD3

E-CAD4
C. Atomic models fit to EM surfaces
E-CAD5

-catenin

FIGURE 30.6 ADHESION BY CADHERINS. A, Electron micrograph of a thin section of a desmosome, colorized to emphasize the plasma
membranes (red) and cadherin extracellular domains (blue). B, Three-dimensional reconstructions of the plasma membrane and cadherin extracel-
lular domains. C, Crystal structure of the C-cadherin extracellular domains fit into electron microscopic reconstructions of intercellular links between
the cells. D, Ribbon diagrams of the crystal structure of a dimer of C-cadherin extracellular domains compared with the book icon for cadherins.
The inset highlights the antiparallel intermolecular interaction of the two CAD1 domains mediated by flexible N-terminal peptides. A conserved
tryptophan fits into a hydrophobic pocket of the partner CAD1 domain, forming the reciprocal interactions. Calcium ions (blue) stabilize interactions
between CAD domains. (AC, From He W, Cowin P, Stokes DL. Untangling desmosomal knots with electron tomography. Science. 2003;302:109
113, copyright the American Association for the Advancement of Science. D, For reference, see PDB file 1L3W and Boggen TJ, Murray J,
Chappuis-Flament S, etal. C-Cadherin ectodomain structure and implications for cell adhesion mechanisms. Science. 2002;296:13081313.)

Wnt
Cadherin
Wnt receptor

-catenin bound
to cadherin
Free -catenin
APC
-catenin Casein
CYTOPLASM bound to kinase 1
APC GSK
E3 ubiquitin
Transcription Tcf ligase SCFTRCP
factors Degradation
of -catenin by
DNA Gene
proteasomes
NUCLEUS expression

FIGURE 30.7 PARTICIPATION OF -CATENIN IN GENE EXPRESSION. Free -catenin is in equilibrium with binding sites on cadherins
and APC (adenomatous polyposis coli) and may also enter the nucleus, where it combines with Tcf/LEF-1 transcription factors. When -catenin
concentrations are low in the nucleus, Tcf/LEF-1 represses gene expression, but the complex of -catenin with Tcf/LEF-1 activates the expression
of genes for cellular growth and differentiation. Synthesis of -catenin is constant, so degradation determines its concentration in cytoplasm:
glycogen synthetase kinase (GSK) and casein kinase-1 phosphorylate -catenin bound to APC, triggering its ubiquitinylation and degradation.
Extracellular Wnt acts through a seven-helix receptor and another receptor to promote gene expression by inhibiting the phosphorylation and
degradation of -catenin.
532 SECTION VIII n Cellular Adhesion and the Extracellular Matrix
Mutations can alter the balance of -catenin synthesis
and turnover. Loss of APCs or mutations of phosphoryla-
tion sites on -catenin result in excess -catenin that
enters the nucleus and stimulates proliferation.
The cadherin expressed from the RET protoonco-
gene signals through its cytoplasmic tyrosine kinase
domain (Fig. 30.5). Point mutations in the segment
between the CAD domains and the plasma membrane
or tyrosine kinase of RET cause constitutive dimerization
of the receptor or activation of the tyrosine kinase
or both. These mutations cause dominantly inherited
cancers of endocrine glands. On the other hand, muta-
tions that disable RET cause Hirschsprung disease.
Autonomic nerves in the wall of the intestines fail to
develop, causing severe dysfunction.
Roles of Cadherins in Organ Formation
Differential expression and regulation of cadherins help
guide organ formation during embryonic development
(Fig. 30.8). Cells with matching cadherins bind together
and exclude cells without those cadherins (or other
appropriate adhesion receptors), although the mecha-
nism is more complicated than differential affinities of
cadherins for each other. For example, cadherins can be
activated or inactivated from inside the cell by signaling
pathways in response to growth factors or other adhe-
sion proteins. In other situations, IgCAMs facilitate the
assembly of cadherins in adhesive junctions.
All cells of early embryos express several different
cadherins, but as soon as the embryo forms three germ
layers, the ectoderm on the outside surface expresses
E-cadherin. In its absence, embryos die. Subsequently,
when ectoderm folds inward to form the neural tube,
those cells switch to expressing N-cadherin. Neurons
use protocadherins to avoid making synapses with
themselves (see below). Later in development, cells
in specialized organs typically express characteristic
A B
Ectoderm
Neural tube
L-CAM (E-cadherin)
Cadherin 6B
N-cadherin
FIGURE 30.8 RESTRICTED EXPRESSION OF CADHERINS
DURING FORMATION OF THE NEURAL TUBE. A, Distribution of
three cadherins before and after the neural tube forms. B, Fluorescent
antibody staining reveals the selective expression of cadherin 6B
(green) and N-cadherin (red) in the neural tube of a developing chick
embryo. (Courtesy M. Takeichi, Kyoto University, Japan.)
cadherins, such as those in osteoblasts (OB-cadherin),
kidney (K-cadherin), and muscle (M-cadherin). A giant-
sized cadherin links sensory stereocilia on the hair cells
in the inner ear. These tip links pull open ion channels
when the stereocilia move in response to sound waves.
Integrin Family of Adhesion Receptors
Integrins are the main cellular receptors for ECM mol-
ecules (Table 30.3), but some integrins bind adhesion
molecules on other cells. Their genes arose early in
metazoan evolution, so they are present in sponges and
corals that branched early in the evolution of animals
(see Fig. 2.8). Fibroblasts and white blood cells use
integrins to adhere to fibronectin and collagen as they
move through the ECM. Integrins bind epithelial and
muscle cells to laminin in the basal lamina, providing the
physical attachments necessary to transmit internal
forces to the matrix and to resist external forces (see Fig.
31.8). These interactions generate signals that control
cell growth and structure. When defects in small blood
vessels need repair, integrins allow platelets to adhere
to basement membrane collagen and to each other via
plasma fibrinogen (see Fig. 30.14). Together, these
interactions are essential for tissue development and
integrity in multicellular organisms. Genetic losses of
integrin function result in several human diseases.
Structure of Integrins
Integrins are heterodimers of two transmembrane poly-
peptides called - and -chains, which both contribute
to ligand-binding specificity (Fig. 30.9). Vertebrate cells
use a combinatorial strategy to establish their integrin
repertoire by selectively expressing a subset of 18 differ-
ent -chains and eight -chains. These chains combine
to form at least 24 different kinds of dimers that bind
different ligands. Alternative messenger RNA (mRNA)
splicing (see Fig. 11.6) adds to the diversity of integrin
isoforms.
The ligand-binding domains of the - and -chains
form a globular head connected to the plasma membrane
by 16-nm legs (Fig. 30.9). More than 25 disulfide bonds
stabilize these domains. All integrin -chains and a subset
of integrin -chains have an I domain (inserted domain)
with a bound divalent cation that interacts with acidic
residues of ligands. All -chains have an N-terminal
-propeller domain similar to a trimeric G protein
-subunit (see Fig. 25.9). Interaction of the -chain
propeller domain with the -chain I domain holds the
integrin dimer together. Single transmembrane segments
anchor both integrin chains to the cell membrane. Short
( 77 residues; = 40 to 60 residues, except 4 = 1000
residues) C-terminal cytoplasmic tails contribute to effi-
cient heterodimer assembly.
The I domains and the -propeller bind at least two
sites on ligands. For instance, integrin 51 binds two
 CHAPTER 30 n Cellular Adhesion 533

TABLE 30.3 Integrin Family of Cell Adhesion Molecules*

Extracellular Intracellular
Examples Structure Ligands Me2+ Ligands Expression Function
Fibronectin receptors 51, others Fibronectin Ca Talin, paxillin Fibroblasts, other cells Cell-matrix adhesion
GPIIb/GPIIIa IIb3 Fibrinogen, von Ca Talin, paxillin Platelets Platelet aggregation
Willebrand factor
Laminin receptor 61, 71 Laminin Yes Talin, paxillin Epithelia, muscle Cell-matrix adhesion
LFA-1 (CD11/CD18) L2 Ig-CAM-1, -2, -3 Mg Talin, paxillin All WBCs WBC/endothelium
adhesion
MAC-1 M2 Ig-CAM-1, Yes Talin, paxillin WBCs except WBC/endothelium
fibrinogen lymphocytes adhesion
Vitronectin receptor V3 Vitronectin, Ca Talin, paxillin Endothelium, smooth
fibronectin muscle, others
VLA-4 21 Collagen, laminin Mg Talin, paxillin WBCs, epithelium, WBC/matrix
endothelium adhesion

CD, cellular differentiation antigen; GP, glycoprotein; ICAM, intercellular adhesion molecule; LFA, lymphocyte functionassociated antigen; Me2+, divalent
cation dependence; VLA, very late antigen; WBC, white blood cell.
*Twenty-four are known.

sites on fibronectin: an arginine-glycine-aspartic acid A -propeller Ig domains

(RGD) sequence on a surface loop of FN-III domain 10
and a secondary site on the adjacent FN-III domain 9 V
Optional
(see Fig. 29.14). Both sites are required for binding, so I domain
simple RGD peptides can compete fibronectin from the
EGF
integrin. Integrin binding sites of some ligands are on I domain domains
Ig-like
separate polypeptide chains. The RDD binding site for
integrin 11 is on three different polypeptide chains of B (Book integrin icon)
the type IV collagen triple helix.
Binding of extracellular ligands (outside-in signaling)
and intracellular ligands (inside-out signaling) influences
the conformation and activity of integrins (Fig. 30.10).
Without bound ligands integrins bend over on closely
C I domain 1, 2, L, M, X
spaced legs held together by interactions of the trans- (~ 1200 aa)
membrane helices. This closed state has a low affinity -propeller 3, 5, 6, IIb, V
for extracellular ligands owing to an occluded binding (~1000 aa)
site. Ligand binding inside or outside the cell favors an
I domain subunit
open state with widely spaced, extended legs holding (~750 aa)
the head above the membrane. In this conformation the EGFs
exposed ligand-binding site has the highest affinity for FIGURE 30.9 INTEGRIN ARCHITECTURE. A, Ribbon diagram of
extracellular ligands. integrin V3 based on a crystal structure of the extracellular domain.
The I domain is inserted into the sequence of an immunoglobulin-
Extracellular Ligands like domain. B, Integrin icon used throughout this book. C, Domain
models of integrin polypeptides. Both -chains and -chains have
Integrins are more promiscuous than most adhesion
single transmembrane segments and cytoplasmic tails that vary in
receptors, as some bind to several protein ligands, and length. All -chains and some -chains have an I domain (red) that
many matrix molecules bind to more than one integrin. binds a divalent cation and participates in ligand binding. The seven
For example, fibronectin binds to at least nine different blades of the -chain -propeller domains are shown in orange. The
integrins, and both laminin and von Willebrand factor -chain I domain, if present, is inserted between the second and third
of the seven blades of its propeller domain. (A, Based on an atomic
bind at least five different integrins. This promiscuity
model. For reference, see PDB file 1JV2 and Xiong JP, Stehle T,
reflects common motifs in the ligands. Approximately Diefenbach B, etal. Crystal structure of the extracellular segment of
one-third of matrix ligands for integrins involve the integrin V3. Science. 2001;294:339345. C, Modified from Kuhn K,
sequence motif RGD or other simple sequences in oth- Eble J. The structural basis of integrin-ligand interactions. Trends Cell
erwise unrelated proteins. Biol. 1994;4:256261.)
Even in the open state (legs apart), integrins generally
have a low affinity for extracellular ligands. For example,
the micromolar Kd for integrin 51 binding fibronectin
534 SECTION VIII n Cellular Adhesion and the Extracellular Matrix

results in rapid association and dissociation, allowing

C. Open state with
B. Open State ligand bound cells to adjust their grip on fibronectin in the matrix as
I-domain Ligand they move through connective tissue. Nonadhesive RGD
-propeller binding
site proteins, such as tenascin (see Fig. 29.16), may modulate
these interactions by competing with fibronectin and
A. Low affinity other ligands for binding integrins.
closed state

-subunit
Intracellular Ligands

-subunit
Talin
FERM
domain
FIGURE 30.10 CONFORMATIONAL STATES OF INTEGRINS.
Drawings based on atomic models derived from crystal structures and
electron microscopy. The bent closed conformation observed in all
crystal structures is inactive. Binding of either an extracellular ligand to
the head or an activated signal transduction protein such as talin to
the cytoplasmic domains can favor the open active state. (Modified
from Xiao T, Takagi J, Coller BS, etal. Structural basis for allostery in
integrins and binding to fibrinogen-mimetic therapeutics. Nature.
2004;432:5967.)
Cytoplasmic tails of integrins interact directly or indi-
rectly with a remarkable variety of signaling and struc-
tural proteins (Fig. 30.11). These interactions are best
understood at focal contacts, specialized sites where
integrins cluster together to transduce transmembrane
signals and link actin filaments across the plasma mem-
brane to the ECM. The adapter proteins talin and vin-
culin link the cytoplasmic domains of -integrins to
actin filaments at the ends of stress fibers. Paxillin links
integrins to signaling proteins, forming a scaffold for Src
family tyrosine kinases (see Fig. 25.3) and focal adhe-
sion kinase (an essential tyrosine kinase).
Several types of integrins associate laterally, in the
plane of the bilayer, with other transmembrane proteins.
The best characterized is CD47 (also called integrin-
associated protein), an IgCAM with five transmembrane

A B GLASS SLIP
ECM
PM
Actin

C EXTRACELLULAR MATRIX

Integrins
FAK (focal
adhesion
kinase)
?
Paxillin
VASP
Crk (SH3 /SH2
Src adaptor)

Csk
Talin Cas (adaptor)
Vinculin

FIGURE 30.11 FOCAL CONTACTS OF EPITHELIAL CELLS WITH THE EXTRACELLULAR MATRIX (ECM). A, Fluorescence micrograph
of parts of two vertebrate tissue culture cells with focal contacts labeled with a fluorescent antibody to phosphotyrosine (orange). Actin filament
stress fibers are stained green with phalloidin. B, Electron micrograph of a thin section of two focal contacts showing fine connections to the
ECM deposited on the surface of the glass coverslip and cross-sections of actin filaments in the cytoplasm. This HeLa (Henrietta Lacks) cell was
grown on a glass coverslip, fixed, and cut perpendicular to the substrate. C, Drawing of the interactions of some of the proteins concentrated
on the cytoplasmic face of the membrane at focal contacts. For clarity, the actin filament interactions (left) are shown separately from some signaling
proteins (right). The rod-shaped dimeric protein talin interacts with the cytoplasmic domains of -integrins and actin filaments. Vinculin interacts
with membrane phospholipids, actin filaments, and talin. An unidentified protein (the question mark) links the adapter protein paxillin to integrins.
Paxillin anchors tyrosine kinases (FAK and Src) and, after phosphorylation, the adapter proteins Crk and Cas. (A, Courtesy K. Burridge, University
of North Carolina, Chapel Hill. B, Courtesy Pamela Maupin, Johns Hopkins University, Baltimore, MD. From Maupin P, Pollard TD. Improved
preservation and staining of HeLa cell actin filaments. J Cell Biol. 1983;96:5162, copyright the Rockefeller University Press. C, For reference,
see Turner C. Paxillin and focal adhesion signaling. Nat Cell Biol. 2000;2:E231E236; and Critchley DR. Focal adhesionsthe cytoskeletal
connection. Curr Opin Cell Biol. 2000;12:133139.)

segments. Binding of the adhesive glycoprotein, throm-
bospondin, to the extracellular Ig-like domain of CD47
generates a transmembrane signal through trimeric
G-proteins that contributes to neutrophil and platelet
activation.
Outside-in Signaling From Integrins
Integrin binding to matrix ligands initiates signals that
modify cellular adhesion, locomotion, and gene expres-
sion, with the responses depending on the particular
integrin and cell. Extracellular ligands stabilize the open
state with wide spacing of the cytoplasmic domains.
Presumably this physical change influences the activities
of signal transduction proteins associated with the cyto-
plasmic domains, but the details are not known. Within
seconds, the cytoplasmic tyrosine kinases shown in Fig.
30.11 phosphorylate several focal adhesion proteins,
including paxillin, tensin, and focal adhesion kinase,
which has a central role in transducing these signals.
Within a minute, some cells raise their cytoplasmic Ca2+
concentration high enough to initiate many calcium-
dependent processes (see Chapter 26).
Over a period of minutes, ligand binding to integrins
also activates Rho-family GTPases that stimulate actin
assembly (see Fig. 33.19) and spreading of the cell on
ligand-coated surfaces. Other Rho-family GTPases drive
contraction of the trailing edge of moving cells (see
Fig. 38.6). Integrins cluster together in small focal
complexes at the leading edge and grow into mature
focal contacts (Fig. 30.11A), also called focal adhesions,
which anchor actin filament stress fibers to the cell
membrane. Contraction of stress fibers applies tension
to the focal contacts, which remain stationary as the
cell advances past them. Rapid rearrangements of the
linker proteins between the integrins and actin serve
as a molecular clutch to transmit forces, even as actin
assemble and disassembles. A Ca2+-mediated signal inac-
tivates obsolete attachments at the rear of the cell.
The adhesiveness of a cell for its substrate (a function
of integrin density on the cell, ligand density on the
substratum, and their affinity) determines the rate of
movement. The maximum rate occurs at intermediate
adhesiveness. Rapid association and dissociation of
integrins on matrix ligands allow cells to rearrange their
hold on the matrix as they move.
After several hours of integrin engagement, activation
of the Ras/mitogen-activated protein kinase pathway
(see Fig. 27.6) turns on expression of selected genes.
These changes in gene expression contribute to cellular
differentiation during development. Integrins allow cells
to include the ECM as a signaling input along with other
stimuli operating through different receptors.
Inside-Out Signaling to Integrins
Cells fine-tune their interactions with matrix molecules
by regulating the activity of cell-surface integrins. For
CHAPTER 30 n Cellular Adhesion 535
example, integrins on white blood cells (Fig. 30.13) and
platelets (Fig. 30.14) require inside-out activation
before they can bind their extracellular ligands. Integrin
activation also regulates cellular interactions during
development. Cytoplasmic proteins, talin and kindlins,
activate integrins by binding the cytoplasmic tail of the
-integrin and separating the two transmembrane
domains. One pathway downstream from seven-helix
receptors uses a membrane-bound GTPase and an
adapter protein to bring together talin and the -integrin.
Some cells can mobilize a reserve pool of integrins
stored in cytoplasmic vesicles within minutes. For
example, chemoattractants stimulate white blood cells
to fuse storage vesicles containing integrins with the
plasma membrane (Fig. 30.13). Both intracellular and
extracellular ligands can cluster integrins in focal com-
plexes and focal adhesions and increase their activities.
Biological Functions of Integrins
With the exception of red blood cells, integrins are
present in the plasma membranes of most animal cells.
Experiments with neutralizing antibodies, genetic dis-
eases, and experimental gene disruptions revealed the
functions of integrin isoforms. For example, many verte-
brate cells express 1 and 3 integrins for adhesion to the
ECM, so integrin antibodies inhibit cell migration and
embryonic development by competing with fibronectin.
Like null mutations in the fibronectin gene (see Fig.
29.14), homozygous disruption of the integrin 4 or 5
genes is lethal during development. Only white blood
cells express 2-integrins, which they use to bind endo-
thelial cells lining the walls of blood vessels.
Other integrins bind adhesion molecules on other
cells. For example, mouse sperm bind integrins on the
egg membrane during fertilization. Integrins cooperate
with adhesion receptors of the IgCAM, mucin, and
selectin families to facilitate the adhesion of white blood
cells to endothelial cells at sites of inflammation (Fig.
30.13). Other cells supplement the functions of integrins
with structurally distinct matrix adhesion proteins, such
as muscle dystroglycans and platelet GPIb-IX-V.
Integrins also participate in the decision of cells to
undergo apoptosis, programmed cell death (see Chapter
46). Normal epithelial cells require anchorage to the
basal lamina by 4-integrins to grow and divide. When
forced to live in suspension or when dissociated from
the matrix by RGD peptides, these cells arrest in the
G1 phase of the cell cycle (see Chapter 41) and eventu-
ally undergo apoptosis. Loss of contact with the basal
lamina may contribute to the terminal differentiation
and death of cells in the upper levels of stratified
epithelia, such as skin (see Figs. 35.6 and 40.1). Epi-
thelial cancers typically lose this integrin-mediated,
anchorage dependence for growth, which is one of
the normal limitations on uncontrolled proliferation in
inappropriate locations.
536 SECTION VIII n Cellular Adhesion and the Extracellular Matrix

Snake venoms contain small, monomeric RGD pro-
teins that inhibit blood clotting by competing with
fibrinogen for binding the integrins that activated plate-
lets use for aggregation. These disintegrins are potential
inhibitors of the pathological thrombosis that contributes
to heart attacks and strokes. Both small-molecule and
antibody antagonists for integrins are now used as clini-
cal treatments for heart attacks and stroke.
Selectin Family of Adhesion Receptors
White blood cells and platelets use three selectin pro-
teins to interact with vascular endothelial cells and each
other. In lymph nodes or at sites of inflammation, selec-
tins snare circulating white blood cells, allowing them
to roll over the surface of endothelial cells and eventually
to exit the blood (Fig. 30.13). Selectins (Table 30.4) also
contribute to adhesion in other systems, including the
initial binding of early mammalian embryos to the wall
of the mothers uterus.
The defining feature of selectins is a calcium-dependent
lectin domain (Fig. 30.12) that binds O-linked sulfated
oligosaccharides containing sialic acid and fucose. The
lectin domain sits at the end of a rod-shaped projection
composed of complement regulatory domains that is
anchored to the plasma membrane by a single transmem-
brane sequence.
Natural ligands for selectins are mucin-like glycopro-
teins expressed on endothelial and white blood cells.
Specific binding to mucins requires selectins to interact
with both the oligosaccharide and mucin protein. The
affinity is low (millimolar Kds), and not highly selective
among oligosaccharides. Interaction with the mucin
protein is less well understood, but one or more sulfated
tyrosine residues on the leukocyte mucin called P-
selectin glycoprotein ligand (PSGL)-1 participate in
binding P-selectin.
Bonds between selectins and their mucin ligands have
high tensile strength (withstanding forces greater than
100 piconewtons [pN]) but form and dissociate rapidly,

TABLE 30.4 Selectin Family (Lec-CAM) of Cell Adhesion Molecules

Examples Structure Extracellular Ligands Me2+ Expression Functions
E-selectin (CD62E, ELAM-1) Lectin-EGF-6CR-1TM L-selectin Ca Endothelium WBC-endothelium
(regulated) adhesion
L-selectin (CD62L, gp90M) Lectin-EGF-2CR-1TM E-selectin, mucins Ca Lymphocytes, WBC-endothelium
other WBCs adhesion
P-selectin (CD62P, Lectin-EGF-9CR-1TM Mucins Ca Endothelium, WBC-endothelium
GMP-140) platelets adhesion
CD, cellular differentiation antigen; CR, complement regulatory domain; EGF, epidermal growth factor; ELAM-1, endothelial-leukocyte adhesion molecule
1; GMP-140, granule membrane protein 140; gp, glycoprotein; Me2+, divalent cation dependence; TM, transmembrane domain; WBC, white blood cell.

A. Leukocyte B. Endothelium

Gly-CAM-1

L-selectin CD34

MAdCAM-1

N-linked oligosaccharide
PSGL-1 O-linked oligosaccharide Complement
regulatory domains
P-selectin

Lectin domain
EGF domain
E-selectin
?

FIGURE 30.12 STRUCTURE OF SELECTINS AND THEIR MUCIN LIGANDS. Domain architecture of selectins and mucins exposed on
the surfaces of leukocytes (A) and endothelial cells (B). Complement regulatory domains of the selectins are shown in red. EGF, epidermal growth
factor; MAdCAM, mucosal addressin cell adhesion molecule; PSGL, P-selectin glycoprotein ligand. (Modified from Rosen SD, Bertozzi CR. The
selectins and their ligands. Curr Opin Cell Biol. 1994;6:663673.)

Selectins
Chemoattractants
1. Attachment
Integrins
2. Rolling
3. Activation
4. Arrest and adhesion
strengthening
5. Transendothelial
Leukocyte migration
Endothelium BLOOD
Basement
membrane
TISSUE
Chemoattractant
source
FIGURE 30.13 MIGRATION OF A NEUTROPHIL FROM THE
BLOOD TO THE CONNECTIVE TISSUE. Endothelial cells exposed
to inflammatory agents like histamine move selectins to their surfaces
and snare mucins on neutrophils flowing in the bloodstream (1). As a
neutrophil rolls along the surface (2), chemotactic factors and engage-
ment of mucins activate their integrins (3), causing the neutrophil
to bind tightly to immunoglobulin cell adhesion molecules (IgCAMs)
on the endothelium (4). The neutrophil then migrates between the
endothelial cells into the connective tissue (5). (For reference, see
Springer T. Traffic signals for lymphocyte and leukocyte emigration: the
multi-step paradigm. Cell. 1994;76:301314.)
on a second time scale. Low forces on these bonds
prolong their lifetimes modestly by altering the confor-
mation of the selectin, whereas high forces promote
dissociation. Consequently, few selectin-mucin bonds
are required to tether white blood cells to the endothe-
lium, whereas the brief lifetime of the bonds allows
blood flow to propel the cells with a rolling motion over
the surface of the endothelium (Fig. 30.13).
Engagement of selectins with mucins stimulates
signals in both cells that activate integrins and promote
adhesion. The process resembles T-cell activation and
includes Src-family kinases and adapter proteins with
tyrosine phosphorylation sites. However, the links from
the cytoplasmic domains of selectins and mucins to the
signaling proteins is not established.
Inflammatory mediators regulate selectins in multiple
ways. Activation of endothelial cells with histamine or
platelets with thrombin causes vesicles storing P-selectin
to fuse with the plasma membrane, exposing the selec-
tins on the cell surface. Various inflammatory agents
stimulate endothelial cells to synthesize E-selectin and
P-selectin. Activation of white blood cells increases the
affinity of L-selectin for mucins and later leads to its
proteolytic release from the cell surface.
Mucins
The extracellular segments of mucins are rich in serine
and threonine, which are heavily modified with acidic
oligosaccharide chains (Fig. 30.12). Because of their
CHAPTER 30 n Cellular Adhesion 537
strong negative charge, these proteins extend like rods
up to 50nm from the cell surface. Mucins on endothelial
cells or white blood cells interact with complementary
selectins on the other cell type. Endothelial mucin CD34
interacts with white blood cell L-selectin, whereas endo-
thelial P-selectin interacts with white blood cell PSGL-1
mucin. These interactions depend on anchoring of the
cytoplasmic domain of PSGL-1 to the actin cytoskeleton.
Other mucins are displayed on the surface of or secreted
by epithelia lining the respiratory and gastrointestinal
tracks.
Other Adhesion Receptors
Table 30.5 lists a variety of adhesion receptors that fall
outside the five main families. See Fig. 25.6 for the CD45
phosphatase, Fig. 31.3 for claudins and Fig. 31.7 for
connexins.
Galactosyltransferase
The enzyme galactosyltransferase is also an adhesion
receptor. This enzyme is a resident protein in the Golgi
apparatus where it glycosylates proteins (see Chapter
21). However, the mRNA for galactosyltransferase has
two alternative initiation sites, one of which adds 13
amino acids to the cytoplasmic, N-terminus of this
transmembrane protein. The longer enzyme moves to
the cell surface rather than being retained in the Golgi
apparatus. On the cell surface, the enzyme can bind
oligosaccharides that terminate in N-acetylglucosamine,
which is found on both cell surface and matrix pro-
teins. The complex of transferase and ligand oligosac-
charide is stable, because the galactose-nucleotide
substrate added to the oligosaccharide in the Golgi
apparatus is not available outside the cell to complete
the reaction.
During fertilization, a surface galactosyltransferase
mediates the initial contact of mouse sperm with the
matrix surrounding the egg (called the zona pellucida).
This association induces secretion of the contents of
the sperm acrosomal vesicle, including an enzyme that
destroys the transferase binding site on the matrix
so that the sperm can proceed through the zona to
fuse with the egg. The enzyme is present on the surface
of many cells that migrate during embryogenesis
and may contribute to their interactions with the
matrix.
Adhesion Receptors With Leucine-Rich
Repeats (GPIb-IX-V)
The platelet receptor for the adhesive glycoprotein
called von Willebrand factor (Fig. 30.14) is a disulfide-
bonded complex of four transmembrane polypeptides:
GPIb, GPIb, GPIX, and GPV. Leucine-rich repeats at
the end of a long stalk bind von Willebrand factor (see
Fig. 28.6 for other receptors with leucine-rich repeats).
538 SECTION VIII n Cellular Adhesion and the Extracellular Matrix

TABLE 30.5 Other Cell Adhesion Molecules

Extracellular Intracellular
Examples Structure Ligands Me2+ Ligands Expression Functions
CD44 Link protein-1TM Hyaluronan Ankyrin Lymphocytes Adhesion to
endothelium
CD44E Link protein-HS/ Fibronectin, No Many epithelial cells Adhesion to
CS-1TM hyaluronan matrix
Claudins Four TM Claudins No ZO-1, ZO-2, Epithelia Tight junctions
ZO-3, cingulin
Connexins Multispan, hexamer Self No ZO-1 Epithelia, muscle, Gap junctions
nerve
Dystroglycans Multisubunit, TM Laminin, agrin Ca Dystrophin Muscle Adhesion, synapse
formation
Galactosyl- Galactose transferase- N-acetylglucosamine No ? Actin Many cells, including Adhesion to cells
transferase 1TM filaments sperm and matrix
Glypican 4HS-GPI anchor Fibronectin, No None Endothelium, smooth Adhesion to
muscle, epithelium matrix
GPIB-IX 7 leucine-rich-1TM von Willebrand Filamin, actin Platelets, Adhesion
factor endothelium
LCA (CD45) 50kD-1TM-tyrosine WBCs Tyrosine
phosphatase phosphatase
Mucins (CD34, Sialylated Selectins No Epithelia, leukocytes Intercellular
CD43) oligosaccharide-1TM adhesion

CD, cellular differentiation antigen; CS, chondroitin sulfate; GPI, glycosylphosphatidylinositol; HS, heparan sulfate; LCA, leukocyte common antigen;
Me2+, divalent cation dependence; TM, transmembrane domain; WBC, white blood cell.

Platelets bind to von Willebrand factor to initiate the
repair of damaged blood vessels. This interaction also
generates an intracellular signal that enhances affinity
of integrin IIb3 for fibrinogen and reorganizes the
cytoskeleton.
Dystroglycan/Sarcoglycan Complex
In muscles, a complex of transmembrane glycoproteins
links a network of dystrophin and actin filaments on the
inside of the plasma membrane to two proteins of the
extracellular basal lamina, 2 laminin and agrin (see
Fig. 39.17). These protein associations stabilize the
muscle plasma membrane from inside and outside,
similar to the actin-spectrin network of red blood
cells (see Fig. 13.11). Genetic defects or deficiencies
in dystrophin, transmembrane linker proteins of the
dystroglycan/sarcoglycan complex, or 2-laminin cause
muscular dystrophy in humans, most likely owing to
the mechanical instability of the membrane, leading to
cellular damage and eventual atrophy of the muscle.
Chapter 39 provides details on their role in muscle func-
tion and disease.
Nonmuscle cells in other tissues express many of
these proteins (or their homologs), where they may
contribute to adhesion to the ECM. Some pathogens
use the dystroglycan complex to bind their cellular
targets. Arenavirus, the cause of Lassa fever, binds
directly to -dystroglycan, and the leprosy bacterium
binds laminin-2.
Examples of Dynamic Adhesion
Adhesion of Leukocytes to Endothelial Cells
Movement of white blood cells from blood into connec-
tive tissue illustrates how cells integrate the activities of
selectins, mucins, integrins, IgCAMs, and chemoattrac-
tant receptors. Infection or inflammation in connective
tissue attracts lymphocytes as well as neutrophils and
monocytes, the main phagocytes circulating in blood
(see Fig. 28.7). Blood cell precursors use a similar mecha-
nism to enter lymphoid organs and bone marrow.
In the absence of inflammation, neutrophils flow over
endothelial cells without binding, because the appropri-
ate pairs of adhesion molecules are not exposed or
activated. Infection or other inflammation in nearby
tissues causes neutrophils to bind to the vascular endo-
thelium and to move out of the blood into the tissue.
Neutrophils adhere to the endothelium in three sequen-
tial but overlapping steps (Fig. 30.13):
1. Locally generated inflammatory molecules, including
histamine (secreted by mast cells), bind to seven-helix
receptors on endothelial cells and stimulate fusion
of cytoplasmic vesicles (called Weibel-Palade bodies)
with the plasma membrane. This exposes P-selectin,
formerly stored in the vesicle membranes, on the
cell surface facing the blood. Selectins bind mucins
that are constitutively exposed on the surface of
neutrophils, tethering them to the surface. The bonds
form and break rapidly, allowing the neutrophil
 CHAPTER 30 n Cellular Adhesion 539

A B C D E
Platelet Damage exposes Activated platelets Activated platelets
basal lamina secrete ADP aggregate over
Endothelium defect
Platelet ADP
Basal lamina binds
ADP

F. Resting platelet G. Activated platelet

Fibrinogen Platelets aggregate

Three independent when fibrinogen
stimuli activate crosslinks
Inactive fibrinogen
receptor platelets
(integrin IIb3) GP1B
(3) ADP activates IIb3 integrin
E T 7-helix receptors (active) binds
EL fibrinogen
AT
PL IIb3 integrin
G (inactive)
TI
N (2) Thrombin activates
vWF
Vesicle ES 7-helix receptors Two pathways
R vWF binds
containing M to collagen activate IIb3
ADP IU GPIb
EL binds
TH to vWF
D
O
IN
A ADP secretion
EN LA
M
T
Proteins not to scale

C L
TA SA
IN BA
(1) Integrin 21
Active collagen receptor binds collagen in
(integrin 21) basal lamina
7-helix ADPreceptor
7-helix thrombin receptor

FIGURE 30.14 PLATELET ACTIVATION AND AGGREGATION AT A DEFECT IN THE ENDOTHELIUM. AD, Steps in platelet activation
and aggregation. E, Electron micrograph of a thin section of a platelet adhering to the basal lamina through a tiny defect in the endothelium.
F, Resting platelets circulate in the blood without interacting with the intact endothelium lining the vessel. G, Platelets are activated in three ways.
(1) Binding of 21 integrins to collagen results in firm adhesion. Where the basal lamina is exposed, von Willebrand factor (vWF) binds the collagen.
(2) Platelet GPIb-IX binds weakly to von Willebrand factor, allowing platelets to adhere to the exposed matrix. (3) Thrombin activates seven-helix
receptors. These interactions stimulate secretion of adenosine diphosphate (ADP), which binds seven-helix receptors and activates the IIb3
integrins; then IIb3 integrins bind dimeric fibrinogen and aggregate platelets together. Platelet proteins are not to scale.

to roll along the surface of the endothelium at Defects in either the weak or strong interactions
rates greater than 10m/s as the blood flow pushes compromise the movement of leukocytes into connec-
them along. tive tissue, increasing the risk of acute and chronic
2. Two signaling pathways (seven-helix receptors for infections. One type of human leukocyte adhesion
chemotactic factors and P-selectin glycoprotein ligand deficiency is caused by a genetic defect in fucose
[PSGL]) activate leukocyte integrins from inside the metabolism that interferes with the synthesis of a car-
cell (Fig. 30.10). Activation of approximately 10% of bohydrate ligand on leukocytes that binds endothelial
the neutrophil integrins increases their affinity for selectins. Cells cannot roll, so they fail to initiate the
their ligand by 200-fold, making the third step emigration process. A genetic deficiency of 2-integrins
possible. causes a second type of leukocyte adhesion deficiency.
3. Activated integrins bind tightly to IgCAMs on the White blood cells that lack 2-integrins roll on the
surface of endothelial cells, immobilizing the leuko- endothelium through the selectin mechanism but do
cyte despite the force of the blood flow. Within 2 not bind tightly enough to migrate out of the circula-
minutes, the leukocyte opens the tight junctions tion. Consequently, these individuals are susceptible to
between endothelial cells (see Chapter 31) and bacterial infections.
squeezes into connective tissue toward the source of On the other hand, neutrophils also generate reactive
the chemoattractant. The leukocyte and endothelial oxygen species that can damage tissues at sites of inflam-
cells interact closely during this passage, because they mation or at sites that are temporarily deprived of
share a self-associating IgCAM called platelet endothe- oxygen. Thus, movement of white blood cells into
lial cell adhesion molecule (PECAM). tissues contributes to damage that occurs when blood
540 SECTION VIII n Cellular Adhesion and the Extracellular Matrix
flow is restored to an ischemic tissue. Therefore adhe-
sion proteins might be targeted therapeutically to miti-
gate damage after heart attacks or severe frostbite.
A similar mechanism and a partially overlapping set
of receptors attract blood monocytes and eosinophils to
sites of inflammation. Once they are in connective tissue,
interactions of monocyte integrins with matrix molecules
trigger the expression of genes required for differentia-
tion into macrophages (see Fig. 28.7).
Lymphocytes (see Fig. 28.9) patrol the body, circu-
lating from the blood through organs to lymphoid
tissues and through the lymphatic circulation back to
the blood. This recirculation requires lymphocytes to
recognize endothelial cells in organs and specific lym-
phoid tissues where they exit from the blood. Lympho-
cytes use L-selectin, three different mucin-like proteins,
and 42 integrins to bind to these target endothelial
cells. Lymphocytes from mice that lack L-selectin
do not roll on endothelial cells or accumulate in
lymph nodes.
Platelet Activation and Adhesion
Platelets aggregate at sites where damage to vascular
endothelial cells exposes the underlying basal lamina
(Fig. 30.14). This process requires the coordinated
activity of a variety of receptors, including integrins,
leucine-rich repeat adhesion proteins, and seven-helix
receptors. These reactions prevent bleeding and bruis-
ing, but inappropriate activation of platelets produces
clots in blood vessels, causing heart attacks and strokes.
To understand the good effects and avert the bad,
investigators have studied platelet activation and adhe-
sion in great detail.
Resting platelets have a low tendency to aggregate,
even though they circulate in a sea of ligands, including
fibrinogen and the adhesive glycoprotein von Willebrand
factor. Multiple mechanisms limit the reactivity of
resting platelets. First, the major integrin, IIb3, has a
low affinity (Kd M) for its plasma ligand, fibrinogen.
Similarly, the GPIb-IX-V complex has a low affinity
for soluble von Willebrand factor. Third, the endothe-
lium masks potential ligands, collagen, and von Wille-
brand factor in the basal lamina. The concentrations
of soluble activators, such as adenosine diphosphate
(ADP) and thrombin, are low under physiological
conditions.
Damage to the endothelium usually initiates platelet
activation by exposing platelets to von Willebrand
factor and collagen in the basal lamina. Under condi-
tions of high shear, GPIb-IX-V interacts strongly with
von Willebrand factor bound to basal lamina collagen.
This interaction transiently tethers platelets to the basal
lamina and favors binding of integrin 21 to collagen.
Exposure to soluble agonists such as ADP or thrombin
also activates platelets and promotes their aggregation.
Within seconds of activation, platelet IIb3 integrins
convert to a high-affinity state (Kd < M) and bind
tightly to fibrinogen. Dimeric fibrinogen links platelets
into aggregates.
Agonists activate platelet IIb3 integrins through
three different pathways:
1. Collagen binding to 21 integrin directly stimulates
platelets to activate IIb3 integrins, secrete ADP, and
synthesize the lipid second messenger thromboxane
A2 (see Fig. 26.9).
2. Damage to blood vessels activates the blood-clotting
proteolytic enzyme thrombin, which binds two related
seven-helix receptors and signals through trimeric
G-proteins (see Fig. 25.9) to activate integrin IIb3.
3. von Willebrand factor binding to the platelet receptor
GPIb-IX-V activates IIb3 integrins.
Two additional mechanisms augment all these
responses. Activated platelets secrete ADP, which binds
two types of seven-helix receptors that amplify the
response to thrombin. Aggregation of platelets by binding
dimeric fibrinogen further stimulates their response to
ADP and thrombin.
Platelet aggregation is disadvantageous in the normal
circulation, so several mechanisms actively inhibit
platelet activation. Endothelial cells produce both
nitric oxide and an eicosanoid, prostacyclin (PGI2),
which inhibit platelet activation (see Fig. 26.9). Nitric
oxide acts through cyclic guanosine monophosphate
(cGMP), and PGI2 acts through cyclic adenosine mono-
phosphate (cAMP; see Fig. 26.1). Antibodies and small
molecule drugs that inhibit IIb3 are used to treat heart
attacks.
The most common human bleeding disorder is
von Willebrand disease, caused by mutations in von
Willebrand factor or its receptor, the GPIb subunit of
GPIb-IX-V. Some mutations reduce the concentration of
the factor in blood or reduce the affinity of the factor for
its receptor. Remarkably, mutations in either the factor
or receptor that increase their affinity for each other
also cause bleeding. These high-affinity interactions
cause platelets to aggregate and be removed from the
blood. Loss-of-function mutations in GPIb cause the
human bleeding disorder called Bernard-Soulier syn-
drome. Individuals with Glanzmann thrombasthenia
bleed abnormally because IIb3 integrin is absent or
defective, and their platelets do not aggregate.
Self-Avoidance in the Nervous System
Surprisingly, neurons use adhesion proteins to avoid
forming synapses with themselves by repelling axons
and dendrites from the same cell (Fig. 30.15). Insects
use a family of DSCAM1 IgCAMs for this self-avoidance.
Alternative splicing of the pre-mRNA in each cell gener-
ates a subset of the many thousands of different
DSCAM1 isoforms. These large IgCAMs have 10 Ig-
domains and six FN-III domains that make homophilic
interactions between three Ig-domains of each protein.

Axon
Cell
body
100 m
FIGURE 30.15 SELF-AVOIDANCE OF NEURITES OF CLASS
IV SENSORY NEURONS OF A DROSOPHILA LARVA. Stack of
fluorescence micrographs ~20 m thick of neurons expressing a
mouse transmembrane protein called CD8 tagged with GFP. False
coloring is used to distinguish the cells. The central neuron is red.
(Courtesy Sujoy Ganguly and Jonathan Howard, Yale University, New
Haven, CT.)
Contacts between neurites with the same isoforms
cause repulsion.
Self-avoidance of vertebrate neurons depends on a
family of 48 cadherins called protocadherins. Alterna-
tive splicing of transcripts from three gene clusters
produces a unique mixture of protocadherins on each
neuron. These protocadherins form random dimers in
the membrane and the extracellular domains bind
homophilically to like protocadherins. When a process
from a neuron contacts another part of itself, extensive
interactions between like protocadherins somehow
create a signal that repels the neurite. Weaker interac-
tions between mixtures of protocadherins on different
cells do not generate a repulsive signal and allow
CHAPTER 30 n Cellular Adhesion 541
N-cadherins and other adhesion proteins to specify
synaptic connections. A point mutation in one proto-
cadherin gene is a common cause of human deafness
and blindness.
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