Review articles

The collagen family members as cell adhesion proteins

Jyrki Heino
Summary The collagen family of extracellular matrix proteins has played a fundamental role in the evolution of multicellular animals. At the present, 28 triple helical proteins have been named as collagens and they can be divided into several subgroups based on their structural and functional properties. In tissues, the cells are anchored to collagenous structures. Often the interaction is indirect and mediated by matrix glycoproteins, but cells also express receptors, which have the ability to directly bind to the triple helical domains in collagens. Some receptors bind to sites that are abundant in all collagens. However, increasing evidence indicates that the coevolution of collagens and cell adhesion mechanisms has given rise to receptors that bind to specific motifs in collagens. These receptors may also recognize the different members of the large collagen family in a selective manner. This review summarizes the present knowledge about the properties of collagen subtypes as cell adhesion proteins. BioEssays 29:10011010, 2007. 2007 Wiley Periodicals, Inc. The vertebrate family of collagens can be divided into eight subfamilies The vertebrate family of collagens contains more than 40 genes. The genes code for collagen a chains, which can form at least 28 different collagen subtypes.(1) Collagens typically contain triple helical domains composed of a chains with repeated glycine-X-Y motifs (X and Y mean any of the 20 amino acids in proteins). Post-translational modifications, e.g. hydroxylation of proline and lysine residues by specific enzymes, are also pronounced features in collagens. Proline is often found in X position and it is often hydroxylated when expressed in Y position. Collagen-type triple helical domains can also be found in twenty other proteins, which are not named as collagens, including an enzyme acetylcholinesterase and subcomponent C1q of complement. To be named as a collagen, a protein must have, in addition to the collagen-like structure, a function as a structural protein of the extracellular matrix. However, some proteins with obvious structural and functional similarities with the collagens, such as ectodysplasin A,(2) have no collagen name or number. The sea sponges are among the most primitive metazoans living today. They are known to have two different collagen subtypes.(3) One resembles the fibril-forming collagens in vertebrates (collagens I, II, III, V, XI, XXIV, XXVII) and the other is non-fibrillar and shares some features with vertebrate basement membrane collagen (collagen IV). The two sponge collagens might be the closest descendants of the original collagen that evolved at about the same time as multicellularity. Other vertebrate collagen subfamilies include fibril-associated collagens with interruptions in triple helix (FACITs; collagens IX, XII, XIV, XVI, XIX, XX, XXI, XXII, XXVI). Some of the FACITs decorate the surfaces of collagen fibrils and mediate their interactions with other matrix proteins. There is no direct evidence about the association of all FACITs to fibrils and some FACITs seem to have other functions. The triple-helical proteins that also have transmembrane domains form an interesting subgroup of the collagens (collagens XIII, XVII, XXIII, XXV).(4) These cell surface collagens participate in the formation of cellmatrix contact sites such as hemidesmosomes and focal adhesions. Furthermore their extracellular parts can be proteolytically shed out from cell surface and the ectodomains can contribute to the remodification of the matrix.(4) Certain collagen subtypes can also form hexagonal networks (collagens VIII and X), anchoring fibrils (collagen VII) or beaded filaments (VI). The latest collagen, type XXVIII, seems to resemble beaded filament forming collagen.(5) A subgroup of collagens, called the multiplexins (collagens XV and XVIII), participates in the stabilization of the basement membrane structures.

Department of Biochemistry and Food Chemistry, University of Turku, Arcanum, Vatselankatu 2, FI-20014 Turku, Finland. E-mail: jyheino@utu.fi DOI 10.1002/bies.20636 Published online in Wiley InterScience (www.interscience.wiley.com).

Abbreviations: DDR, discoidin domain receptor; FACIT, fibril-associated collagen with interruptions in triple helix; GP , glycoprotein; LAIR, leukocyte-associated immuglobulin-like receptor; MLBR, major ligandbinding region; MMP , matrix metalloproteinase; NC, non-collagenous; uPARAP , urokinase-type plasminogen activator receptor associated protein; VLA, very late activation antigen.

Cells have receptors that can directly bind to collagens Collagens are found in all metazoans and they most probably have contributed to the early evolution of multicellular animals. Surprisingly, the receptors that anchor cells directly to

BioEssays 29:10011010, 2007 Wiley Periodicals, Inc.

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collagens seem to be much younger in terms of their evolutionary history.(6) Integrins are the largest family of cell adhesion receptors.(7) Integrin-type receptors are composed of one a and one b subunittwo transmembrane proteins that are noncovalently linked together. Integrin a and b subunits are evolutionarily unrelated. Based on phylogenic reconstruction, integrin a subunits seem to segregate into five different subgroups, four of which are present in vertebrates.(6) The largest subgroup in vertebrates is formed by a subunits that have a special ligandbinding inserted domain, aI domainalso called aA domain, based on its similarity to von Willenbrand factor A domain. Despite the fact that in the human integrin family nine out of eighteen a subunits have aI domains, this subgroup of the integrins is not present in insects or worms.(6) It was recently shown that urochordate genomes carry genes for aI domain integrins, but the functional relationship between urochordate and vertebrate aI domain integrins is not clear.(6,8) Four integrins containing aI domain subunits a1, a2, a10, and a11, are considered to form the subgroup of the vertebrate collagen receptor integrins. The collagen-binding aI domains have structural details that make them different when compared to other vertebrate aI domain integrins, which seem to function mainly in immunological processes. All collagen receptor a subunits form a heterodimer with a common b1 subunit. Integrin a1b1 (other name: very late activation antigen 1, VLA-1) is expressed in many mesenchymal cell types(9) and it seems to be important for the action of many inflammatory cells, including T lymphocytes. Integrin a2b1 (other names: VLA-2, GPIa/IIa) is abundant on many epithelial cell types and platelets, but it is also expressed on mesenchymal cells.(10) Thus, many mesenchymal cells can express both collagen receptors at the same time. Interestingly, a1b1 and a2b1 integrins seem to have opposite effects on many signalling pathways.(1113) Thus, cellular response to collagens may be dependent on the dominance of another one of the receptors. Integrins a10b1 and a11b1 have more limited tissue distribution. Integrin a10b1 seems to be specific to cartilage(14) and a11b1 is expressed in many mesenchymal tissues during development.(15) Very little is known about the signalling function of these integrins. Other members of the integrin family have also been reported to bind to collagens. Integrin a3b1 may be an assisting collagen receptor(16) and the aI domains of some of the leukocyte integrins, e.g. aXb2 integrin, may bind to collagens.(17) Discoidin domain receptors 1 and 2 (DDR1 and DDR2) are two closely related receptor tyrosine kinases that are known to bind to the collagens(18,19) and act in an integrin-independent manner.(20). It has been proposed that the DDRs are collagen sensors rather than anchoring cell adhesion receptors. Unlike the collagen receptor integrins, DDR-like genes have been found in insects, worms and sponges.(21) Thus it is possible

that invertebrates also have specific receptors for collagens. However, it is not known whether the protein products of the DDR-like genes can recognize collagens. In man and mouse, DDRs are widely expressed in different tissues. DDR1 is considered to be mainly an epithelial and DDR2 mainly a mesenchymal protein.(22) DDRs are autophosphorylated after ligand binding and their signals regulate cell proliferation, differentiation and matrix modulation. Unlike most other receptor tyrosine kinases, the DDRs are phosphorylated hours rather than seconds after ligand binding. Many other plasma membrane proteins have been reported to bind to collagens. Glycoprotein VI (GPVI, p62) is a member of the paired immunoglobulin-like receptor family.(23) It is an important collagen receptor on platelets. Ligand binding to GPVI activates a series of signalling events regulating the activity of other receptors and the function of platelets during thrombosis. CD36 (other names GPIV and GPIIIb) is another platelet receptor binding to collagen I.(24) However, platelets deficient for CD36 show normal response to collagens,(25) challenging its biological role as a collagen receptor. Leukocyte-associated immunoglobulin-like receptor-1 (LAIR-1) is a member of the immunoglobulin superfamily. It is expressed on the peripheral blood mononuclear cells. Antibody-induced clustering of LAIR-1 inhibits the functions of natural killer cells, effector T cells, B cells and dendritic cells. Recently, LAIR-1 was shown to be a high avidity collagen receptor.(26) A member of the macrophage mannose receptor family of endocytic transmembrane glycoproteins, namely uPARAP (urokinase-type plasminogen activator receptor associated protein)/Endo180 seems to play a crucial role in the cellular uptake and lysosomal degradation of collagen.(27) Annexin A5, also called as annexin V or anchorin II, was originally described as a potential receptor for collagen II(28) and was later shown to bind to collagen X as well.(29) Structurally annexin A5 resembles the other members of the annexin family, containing four repeats formed by five a helices and symmetrically grouped around a central pore. Today it is known that annexin A5 is an intracellular protein that can also be secreted. Phosphatidylserine mediates high-avidity binding of annexin 5A on the outer surface of plasma membrane.(30) By binding to collagens, annexin 5A may regulate the influx of Ca2 into matrix vesicles.(29) Annexin 5A is expressed in cartilage and in bone suggesting a role during skeletal development. Proteoglycans can bind to collagens via their glycosaminoglycan chains. CD44 is a class I transmembrane protein expressed in many different splice forms.(31) Some CD44 isoforms also contain glycosaminoglycans. CD44 is a receptor for extracellular matrix hyaluronan molecules. It is abundantly expressed on most vertebrate cell types. CD44 has also been described to bind to collagens I and VI. Interaction of CD44 with collagen I may be dependent on the chondroitin sulphate chains.(32) Cell surface heparan sulphate proteoglycans,

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syndecans and glypicans, can also act as extracellular matrix receptors. Syndecans are transmembrane proteins, whereas glypicans are anchored via glycosylphosphatidylinositol. Syndecan-1 has been described to bind to collagen I(33) and it seems to share this property with syndecan-2 and syndecan4, as well as with glypican-1.(34)

Table 1. Recognition motifs in collagens. Bold letters indicate the critical amino acid residues.
Reference GFOGER site (three collagen a1(I)-like peptides) -GPOGFOGERGPOtrailing strand -GPOGFOGERGPOmiddle strand -GPOGFOGERGPOleading strand A binding site for a1b1 integrin in collagen IV -GAKGRAGFOGLOcollagen a2(IV)-like peptide -GPOGDQGPOGIOcollagen a1(IV)-like peptide -GPOGDQGPOGIOcollagen a1(IV)-like peptide 38

Recognition sites in triple helical and denatured collagens Receptor-binding sites in collagens can be divided into at least four different categories (Fig. 1). First, there are specific motifs inside the triple helical areas that can be recognized by collagen receptors. These motifs represent highly specialized interaction mechanisms and we can speculate that their evolution has required concomitant changes in both receptors and ligands. A second category of receptor-binding sites includes relatively common sequences in the triple helix, such as GPO (O is hydroxyproline). We can presume that, in this case, some receptors have evolved to bind to sites that have already existed in collagens. The third category contains cryptic binding sites in collagens that can be recognized by receptors only after the denaturation of the collagens and the fourth category carries specific receptor-binding sites in noncollagenous (NC) domains, such as NC1 domain. A good example of a specialized collagen-binding mechanism is the GFOGER motif. Triple helices formed by peptides that contain the GFOGER or certain other similar sequences are high-affinity ligands for a2b1 integrin.(35,36) The same motif can also be recognized by the other collagen receptor integrins.(37) The crystal structure of synthetic triple helical GFOGERpeptide in complex with a2I domain has revealed the atomic details of integrincollagen interaction(38) (Table 1). More recently other motifs have been described in collagens. An intensive analysis of collagen III has shown that a2b1 integrin binds with high affinity to motifs such as GROGER and

42

Figure 1. Different receptor-binding sites in collagen molecules. A schematic presentation of possible receptor-binding sites in a collagen molecule. Receptors can bind to both triple helical (collagenous) and non-triple helical (non-collagenous) areas in the molecule.

GLOGEN.(39) Similar motifs are also found in other than fibrilforming collagens suggesting their importance especially in collagens that are homotrimers while, in collagens composed of two or three different a chains, the binding mechanism might be more complex. The binding mechanism of a1b1 to one motif in collagen IV has been analyzed in detail and it seems to be very different when compared to the binding of a2b1 to fibril-forming collagens. The critical binding motif in collagen IV is formed by one arginine (arg461 in a2(IV)) and two aspartic acid residues (asp461 in a1(IV)), all located in different a chains in the triple helix (Table 1; (40,41,42)). The different mechanism of a1b1 binding when compared to a2b1 binding has also been stressed by observations indicating that proline hydroxylation in collagen is more critical for a1b1.(43) It is not known how many different collagen-recognition mechanisms the collagen receptor integrins have. Our observations have proposed that recognition of collagen IX by collagen-binding integrin aI domains may differ from the mechanisms described above.(44) Similarly, the integrin-binding site in triple helical domain of collagen XVI might represent a previously unknown recognition motif.(45) So far the research has been focused on the high affinity integrin-binding sites. However, it is probable that numerous low affinity sites are present in collagens as well. Many collagens seem to have more than one integrin-binding motif and it is possible to speculate that the clusters of recognition motifs may have interesting biological functions. For example, cells may play a central role in the construction of the complex architecture of the extracellular matrix. In theory, the systematically located binding motifs in collagens could form the basis for the cellular guidance. The exact binding motifs in collagen for DDR1 and DDR2 are unknown. DDR2 seems to bind into the triple helical area of collagen II, more precisely into D2 period.(46) Proline hydroxylation seems to be critical for DDR binding.(43) The basic triple helical collagenous structure can be recognized by some receptors. GPVI can bind to peptides containing repeated GPO motifs and GPVI-deficient platelets

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show no response to GPO peptides.(47) GPO motifs are abundant in all collagens and little is known whether GPVI can show selective specificity to some of the collagen subtypes. Despite the fact that GPVI seems to bind to the basic collagenous triple helix, it is possible that the number and order of the GPO motifs regulate the binding specificity. Indeed, the smallest motif that is recognized by GPVI contains two GPO triplets.(48) LAIR-1 also seems to recognize the repeated GPO sequences in the collagens.(26) Neither collagen receptor integrins nor DDRs(18) can recognize heat-denatured collagen, gelatin. Many collagens, e.g. collagen I, contain cryptic RGD motifs, typically recognized by another subset of receptors, namely integrin type fibronectin/vitronectin receptors. Indeed, after heat denaturation of collagen I, a5b1 integrin and aV integrins can bind to it.(49) Importantly, proteolytic degradation of fibril-forming collagens by matrix metalloproteinases (MMPs) may also unveil the hidden RGD motifs. The Col15 domain of collagen XVII contains 12 KGD motifs (four in each a chain). KGD motifs can be recognized by the RGD-binding integrins, but with different affinity. Migrating cells may be able to use the denatured ectodomain of collagen XVII as a migration substrate in an aV integrin-dependent manner.(50) Thus, the cryptic motifs in collagens may be important for collagen turnover and removal of degraded and denatured collagen. Furthermore, after proteolysis collagen fragments may be temporarily integrated into matrix, in which they can provide new adhesion sites for migrating cells. Non-collagenous domains in collagens form a separate group of integrin ligands, which may have interesting biological functions. Integrin a2b1 has been reported to bind to NC1 domains in collagens VII and X,(51) whereas NC1 domains in collagens IV and XVIII are recognized by other integrins. NC1 domain in collagen XVIII is better known as endostatin, a protein with potent anti-angiogenic properties. Its function might be at least partially dependent on its ability to bind to a5b1, aVb3 and aVb5 integrins on endothelial cells.(52,53,54) Six different a subunits, namely a1(IV) a6(IV), may participate in the formation of collagen IV molecules. At least three different collagen IV-derived NC1 domains can inhibit angiogenesis.(55) Aresten is NC1 domain derived from a1(IV) and it is a ligand for a1b1 integrin.(56) Canstatin is from a2(IV) collagen and tumstatin from a3(IV) collagen. They are both ligands for aV integrins.(54,57) a3NC1 domain can also be recognized a3b1 integrin.(58) Thus, receptor-binding sites in collagen NC1 domains seem to be important for cell regulation, especially during angiogenesis.

Distinct recognition of collagen subtypes by adhesion receptors The integrin-type collagen receptors cannot bind to the basic triple helical GPO sequence,(59) but they require specific motifs formed by residues in two or all three a chains (Table 1). Thus, it is probable that the collagen subtypes are different when their properties are compared as cell adhesion proteins. Fig. 2 summarizes the known integrin-type ligands for different collagen subtypes. It is also possible that there are collagens that contain no adhesion motifs for integrins. Indeed, the largest collagenous domain (Col15) in collagen XVII cannot be bound by the collagen receptor integrins.(50) Another similar collagen might be collagen XIV (undulin), a FACITexpressed in soft connective tissues and in cartilage.(60,61) The lack of binding motifs might be essential for the function of these collagens. Collagen IX, a prototype FACIT expressed in cartilage and eye, seems to be a high-avidity ligand for all four collagen receptor integrins.(44) It may have a special role in mediating cell adhesion to fibrils formed by cartilage collagens. Based on collagens IX and XIV, it is possible to speculate that some of the FACITs, e.g. collagen IX, strongly support cell adhesion to fibrils, whereas some others might be anti-adhesion proteins, e.g. collagen XIV, for integrin-type receptors. A third FACIT, collagen XVI, can be recognized by a1b1 integrin and, with lower avidity, by a2b1 integrins.(45) Collagen XVI mediates the anchoring of skin microfibrils to basement membranes rather than interacts with large D-banded fibrils.(62) In published reports, fibril-forming collagens have been high-avidity ligands for a2b1 integrin.(6365) Integrin a2b1 can also bind to fibrils formed by collagen I.(66) The avidity of a2b1 to fibrils might be smaller than to monomeric collagen I,(66) but a2b1 can mediate cell adhesion and migration inside a matrix formed by collagen fibrils. Integrin a2b1 is also a weak receptor for many other collagen subtypes, e.g. collagens IV, VI and XIII.(6365,67) Interestingly, these collagens, especially the basement membrane collagen IV, are high-avidity ligands for a1b1 integrin.(6365) Integrin a1b1 is a low-avidity receptor for fibril-forming collagens IIII and V(6365) and it has been reported to recognize collagens VIII and IX.(44,68) Binding specificities of integrins a10b1 and a11b1 are less well known.(37,65) The analyses based on recombinant aI domains and transfected cells indicate that they are less selective between fibril-forming and other collagen subtypes than a1b1 or a2b1.(37,65) However, a10b1 might resemble more a1b1 than a2b1, whereas a11b1 might prefer fibrilforming collagens in the same way as a2b1.(37,65)

Figure 2. Collagen subfamilies and their integrin-type receptors. The figure lists the existing information about integrin-type receptors for different collagen subtypes, representing eight collagen subfamilies. The basic domain structure of the collagen subtypes has been drawn on the left and, on the right, there is a list of the known receptors. In some cases, the low-avidity receptors are in parentheses.

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The activation stage of the collagen receptors can be regulated. Their ligand-binding aI domains may be in at least two different active conformations.(38) Most information about the binding specificities of recombinant aI domains is based on proteins that are in a closed conformation. The a2I domains, which harbour a specific gain-of-function mutation, favor a high-affinity open conformation. In the active conformation, a2I domain can recognize new binding motifs,(69) and it is likely that binding specificity of all collagen receptor aI domains changes in the activated stage. Our unpublished results propose that selectivity between different collagen subtypes is reduced when the avidity of ligand binding increases after integrin activation. Quite little is known about selective binding of other collagen receptors to collagen subtypes. DDR1 and DDR2 seem to differ in their binding specificity in the way that DDR1 can be activated by both fibril-forming and non-fibril-forming collagens (collagens IVI, VIII), while DDR2 can be activated by fibril-forming collagens (I, II, III and V), only.(19,20) DDR2 seems to be a higher avidity receptor for fibril-forming collagen II than DDR1.(46) The list of cellular receptors for each collagen subtype is far from being complete. Still it can be concluded that some receptors, namely a2b1 integrin, DDR2, and maybe also a11b1 integrin, seem to prefer fibril-forming collagens. Interestingly, both integrin and DDR families have members that have evolved to this direction and cell adhesion to fibrilforming collagens might serve a different purpose when compared to binding to other collagen subtypes. What then makes cell adhesion to fibril-forming collagens or collagen fibrils so special? One reason may be that these collagens are not so often available for cell adhesion receptors. Despite the fact that extracellular matrix is build of collagen fibrils, the fibrils are usually covered with other proteins and proteoglycans making the collagenous recognition motifs unreachable for cellular receptors. The presence of available binding sites may be a sign for the cells about active collagen synthesis or tissue degradation. It has also been suggested that collagen receptors participate in the organisation of collagen monomers into fibrils. In in vitro models, a2b1 and a11b1 integrins promote fibril formation,(70) whereas DDR2 may inhibit it.(71) However, the mechanism of receptor guided fibrillogenesis should also be confirmed in animal models before final conclusions are made. Biological role of direct cell adhesion to collagen Analysis of rare human diseases and manipulation of the mouse genome have given insight to the in vivo function of numerous collagen subtypes.(1) However, the data have a limited value only when the biological significance of direct cellcollagen interaction is estimated. A significant decrease

in the expression of a specific collagen subtype or its complete deficiency may also lead to fundamental changes in tissue architecture, making it impossible to segregate the effects due to the lack of a specific cell adhesion site. Mutations or polymorphisms in collagens may increase the risk of common diseases, such as osteoporosis, osteoarthrosis or arterial aneurysms. It is an interesting possibility that some of these changes affect the cell adhesion motifs in collagens and the consequences reflect the effects of missing collagen receptor functions. Indeed, the first evidence supporting this hypothesis recently came from the comprehensive analysis of lethal mutations in human osteogenesis imperfecta. In collagen I, two exclusively lethal regions seem to align with the major ligand-binding regions (MLBR).(72) Interestingly MLBR2 is known to contain high-affinity binding sites for a1b1 and a2b1 integrins.(73) Mice deficient for a1 integrin subunit have been analyzed for more than ten years(74) and there is an increasing number of reports pointing to specific defects in the immunological system and connective tissues. Integrin a1b1 seems to be important for the proliferation of mesenchymal stem cells. In the absence of a1b1, bone fracture healing is affected,(75) mice develop accelerated ageing-dependent osteoarthritis(76) and their dermis is hypocellular.(77) In addition, a1b1 integrin seems to be a negative feed-back regulator of collagen synthesis.(78) However, the lack of a1b1 seems not to generate skin fibrosis, since the increased matrix turnover rate, due to activated expression of MMPs, compensates the enhanced collagen synthesis.(78) Importantly, a1-deficient mice are more sensitive to induced fibrosis, such as adriamycin-related glomerulosclerosis.(79) Increased MMP expression has also been connected to the impaired tumor-related angiogenesis(80) and osteoarthritis(76) detected in a1 knock-outs. The requirement of a1b1, together with a2b1, for proper angiogenesis has also been confirmed using function-blocking antibodies in the inhibition of tumor angiogenesis in mice.(81) In the immune system, a1b1 is needed for monocyte,(82,83) B lymphocyte and effector/memory T (CD8) lymphocyte functions.(84) Integrin a2b1 is a very abundant collagen receptor expressed in a large variety of different tissues. Therefore it is surprising that the lack of a2 subunit in mouse mainly seems to affect platelet(8587) and mast cell functions.(88) In a2-deficient mice, the branching complexity in mammary glands also seems to be affected,(86) but, for example, re-epithelialisation rate of wounds seems to be normal,(86,89,90) despite the fact that, in human wounds, the migrating keratinocytes have highexpression levels of a2b1 integrin.(91) In the murine cutaneous wound model, neoangiogenesis is enhanced in a2b1 null animals.(89,90) Integrin a10b1 is abundant in cartilage, but its absence has only a small effect on growth plates.(92) Integrin a11 knockout mice are smaller than their wild-type littermates. This is, at least partially, due to severely defective incisors and

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consequent malnutrition,(93) whereas no developmental defects in the skeletal system have been reported. Given the fact that integrin-type collagen receptors have evolved quite recently, it is understandable that they seem not to be critical for embryonic development and the described phenotypes of the knock-out mice are not life-threatening. The phenotypes of DDR1 and DDR2 knockouts have been described. DDR1-deficient mouse are smaller than their littermates(94) and DDR1-null females have defects in the development of mammary glands.(94) In addition, DDR1deficient mice have shown localized subepithelial thickening of basement membranes and proteinuria.(95) DDR2 knockouts exhibit dwarfism and shortening of long bones.(96) In general, DDR2 seems to be important for the proliferation of mesenchymal cells, which has also been seen during wound healing.(96) The phenotypes of collagen receptor knockouts stress the role of collagen receptors in selected biological processes (Table 2). Firstly, both integrins and DDRs participate in the regulation of skeletal development and homeostasis.(93,96) Collagen receptors may also have essential roles during the differentiation of specific cell types, e.g. in breast glands.(87,94) These functions are probably related to receptor signalling and activation of pathways that promote cell growth and survival. Secondly, platelet binding to collagen is a critical phenomenon and is mediated by direct (integrin a2b1 and GPVI)(23,8688) as well as indirect (von Willenbrand factor) mechanisms, which may partially compensate each other. Thirdly, collagen receptors can regulate collagen turnover rate. A typical function of many collagen receptors, including both integrins and DDRs, is to activate the expression of MMPs,(11,18,97) such as collagenases. Collagen receptors also regulate collagen synthesis(11,12,78) and uptake.(27) Finally, collagen receptors seem to participate in many ways in the regulation of immunity and inflammatory response. Inflammatory cells may use their collagen receptors in migration inside the tissues. However,

the role of collagen receptors may be more complex. For example, a2b1 integrin is abundantly expressed on basal keratinocytes, but seems not to participate in cell anchorage or migration. It has been proposed that the task of a2b1 is to react to tissue injury by activation of collagenase expression contributing to innate immunity.(98) Furthermore, some functions of the collagen receptors may be rationalized by the fact that certain collagens act as autoantigens in diseases, such as rheumatoid arthritis and bullous pemphigoid. Therefore, after tissue injury or inflammatory response, collagen molecules and their triple helical fragments must be effectively cleaned out. Collagen binding to its receptor can trigger fast internalization or phagocytosis of the ligand.(99,100) In addition, collagen binding to LAIR-1 suppresses immune responses.(26) A classic example of a collagenous autoantigen is collagen XVII, also called bullous pemphigoid antigen 180, in epidermis. LAIR-1 is present in skin, but collagen can still sometimes escape the immunological tolerance. Interestingly, collagen XVII seems not to be recognized by a2b1 integrin,(50) which is an abundant collagen receptor in epidermis. Could this property of collagen XVII increase the risk of autoimmune response? Based on the present data, it is not possible to link any of the receptor knockout phenotypes to the interaction of the receptor with any single collagen subtype. Similarly, the consequences of the deficiency of any collagen subtype cannot be connected to collagens role as a cell adhesion protein. More information is needed about the molecular architecture of different tissues and the exact cell adhesion motifs in collagen subtypes. Based on this information, it could be possible to create transgenic animals that harbour targeted mutations in receptor-binding motifs of collagens. Conclusion Cells have evolved to recognize various members of the collagen family with surface receptors. Special receptors have

Table 2. Participation of the collagen receptors in the biological functions

Receptors Platelet activation Inammation and immunity Angiogenesis Cancer Wound healing Proliferation of mesenchymal cells Matrix remodelling Collagen synthesis MMP expression Collagen endocytosis/phagocytosis Development Mammary gland Skeleton Periodontal ligament a2b1, GPVI a1b1, a2b1, LAIR-1 a1b1, a2b1 a2b1 a1b1, DDR2 a1b1, a2b1 a2b1, DDR2 a2b1, Endo180 a2b1, DDR1 a10b1, DDR2 a11b1 References 21,86,87,88 26,82,83,84,88 80,81 89,90 75,76,77,96 11,12,78 11,18,97 27,99,100 86,94 92,96 93

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the ability to recognize triple helical domains in collagens. In addition, some receptors bind to non-collagenous domains, e.g. NC1 domains. Finally, a different subset of receptors binds to cryptic sites, such as RGD motifs, in collagen a chain after denaturation of the triple helix (Fig. 1). Structurally different receptors, such as members of integrin, receptor tyrosine kinase and immunoglobulin-like superfamilies, can mediate cell adhesion to collagenous domains. Since the receptors often recognize specific motifs in collagens, it is reasonable to suppose that the 28 members of the collagen superfamily have individual properties as cell adhesion proteins. Based on the present information about the receptor interactions of collagen subtypes (Fig. 2) some conclusions and speculations can be made. Most importantly, some receptors have clearly evolved to favor fibril-forming collagens over other collagen subtypes. It can be hypothesized that adhesion to fibrillar collagen leads to specialized cellular responses or that fibrillogenesis itself requires the participation of cell surface receptors. Another general observation is that the integrin-type collagen receptors cannot recognize all collagen subtypes. Thus, it is possible that both adhesionpromoting and anti-adhesive collagen types exist. Generation of collagen-receptor-deficient mice have helped to understand the essence of cell adhesion to collagen. In general the knockout phenotypes are not very dramatic, although some receptors are clearly important for normal development of bone, cartilage or breast tissue. In addition, collagen receptors seem to participate in the regulation of connective tissue turnover and platelet function. However, abundant expression of collagen receptors in many tissues suggests that all their functions have not yet been found. The participation of collagen receptors in adhesion and migration of inflammatory cells have been known for some time, but new observations connect the receptor function to innate immunity and immunological tolerance to collagen. Detailed information about receptor-binding capabilities and mechanisms of different collagen subtypes will help us to understand the orchestration of tissue architecture. Furthermore, genetic variation in both receptors and collagens may contribute to the pathogenesis of common diseases, such as osteoarthrosis and vascular thrombosis. Acknowledgments The author wants to thank Mr. Timo Kattelus for technical assistance in the drawing of the figures. References
1. Myllyharju J, Kivirikko KI. 2004. Collagens, modifying enzymes and their mutations in humans, flies and worms. Trends Genet 20:3343. 2. Ezer S, Bayes M, Elomaa O, Schlessinger D, Kere J. 1999. Ectodysplasin is a collagenous trimeric type II membrane protein with a tumor necrosis factor-like domain and co-localizes with cytoskeletal structures at lateral and apical surfaces of cells. Hum Mol Genet 8:20792086.

3. Exposito JY, Garrone R. 1990. Characterization of a fibrillar collagen gene in sponges reveals the early evolutionary appearance of two collagen gene families. Proc Natl Acad Sci USA 87:66696673. 4. Franzke CW, Bruckner P, Bruckner-Tuderman L. 2005. Collagenous transmembrane proteins: recent insights into biology and pathology. J Biol Chem 280:40054008. 5. Veit G, Kobbe B, Keene DR, Paulsson M, Koch M, et al. 2006. Collagen XXVIII, a novel von Willebrand factor A domain-containing protein with many imperfections in the collagenous domain. J Biol Chem 281: 34943504. 6. Huhtala M, Heino J, Casciari D, de Luice A, Johnson MS. 2005. Integrin evolution: insights from ascidian and teleost fish genomes. Matrix Biol 24:8395. 7. Hynes RO. 2002. Integrins: bidirectional, allosteric signaling machines. Cell 110:673687. 8. Ewan R, Huxley-Jones J, Mould AP, Humphries MJ, Robertson DL, et al. 2005. The integrins of the urochordate Ciona intestinalis provide novel insights into the molecular evolution of the vertebrate integrin family. BMC Evol Biol 5:31. 9. Voigt S, Gossrau R, Baum O, Loster K, Hofmann W, et al. 1995. Distribution and quantification of alpha 1-integrin subunit in rat organs. Histochem J 27:123132. 10. Zutter MM, Santoro SA. 1990. Widespread histologic distribution of the alpha 2 beta 1 integrin cell-surface collagen receptor. Am J Pathol 137:113120. ha ri VM, et al. 11. Riikonen T, Westermarck J, Koivisto L, Broberg A, Ka 1995. a2b1 integrin is a positive regulator of collagenase (MMP-1) and collagen a1(I) gene expression. J Biol Chem 270:1354813552. ha ri VM, et al. 12. Ivaska J, Reunanen H, Westermarck J, Koivisto L, Ka 1999. Integrin a2b1 mediates isoform specific activation of p38 and upregulation of collagen gene transcription by a mechanism involving the a2 cytoplasmic tail. J Cell Biol 147:401416. ha ri VM, et al. 13. Ivaska J, Nissinen L, Immonen N, Eriksson JE, Ka 2002. Integrin a2b1 promotes activation of protein phosphatase 2A and dephosphorylation of Akt and GSK3b. Mol Cell Biol 22:1352 1359. 14. Camper L, Holmvall K, Wangnerud C, Aszodi A, Lundgren-Akerlund E. 2001. Distribution of the collagen-binding integrin alpha10beta1 during mouse development. Cell Tissue Res 306:107116. 15. Tiger CF, Fougerousse F, Grundstrom G, Velling T, Gullberg D. 2001. alpha11beta1 integrin is a receptor for interstitial collagens involved in cell migration and collagen reorganization on mesenchymal nonmuscle cells. Dev Biol 237:116129. 16. DiPersio CM, Shah S, Hynes RO. 1995. a3Ab1 integrin localizes to focal contacts in response to diverse extracellular matrix proteins. J Cell Sci 108:23212336. 17. Garnotel R, Rittie L, Poitevin S, Monboisse JC, Nguyen P, et al. 2000. Human blood monocytes interact with type I collagen through axb2 integrin (CD11c-CD18, gp150-95). J Immunol 164:59285934. 18. Vogel W, Gish GD, Alves F, Pawson T. 1997. The discoidin domain receptor tyrosine kinases are activated by collagen. Mol Cell 1: 1323. 19. Shrivastava A, Radziejewski C, Campbell E, Kovac L, McGlynn M, et al. 1997. An orphan receptor tyrosine kinase family whose members serve as nonintegrin collagen receptors. Mol Cell 1:2534. 20. Vogel W, Brakebusch C, Fassler R, Alves F, Ruggiero F, et al. 2000. Discoidin domain receptor 1 is activated independently of b(1) integrin. J Biol Chem 275:57795784. 21. Vogel WF, Abdulhussein R, Ford CE. 2006. Sensing extracellular matrix: an update on discoidin domain receptor function. Cell Signal 18:11081116. 22. Alves F, Vogel W, Mossie K, Millauer B, Hofler H, et al. 1995. Distinct structural characteristics of discoidin I subfamily receptor tyrosine kinases and complementary expression in human cancer. Oncogene 10:609618. 23. Moroi M, Jung SM. 2004. Platelet glycoprotein VI: its structure and function. Thromb Res 114:221233. 24. Tandon NN, Kralisz U, Jamieson GA. 1989. Identification of glycoprotein IV (CD36) as a primary receptor for platelet-collagen adhesion. J Biol Chem 264:75767583.

1008

BioEssays 29.10

Review articles

25. Saelman EU, Kehrel B, Hese KM, de Groot PG, Sixma JJ, et al. 1994. Platelet adhesion to collagen and endothelial cell matrix under flow conditions is not dependent on platelet glycoprotein IV. Blood 83:32403244. 26. Lebbink RJ, de Ruiter T, Adelmeijer J, Brenkman AB, van Helvoort JM, et al. 2006. Collagens are functional, high affinity ligands for the inhibitory immune receptor LAIR-1. J Exp Med 203:14191425. 27. Curino AC, Engelholm LH, Yamada SS, Holmbeck K, Lund LR, et al. 2005. Intracellular collagen degradation mediated by uPARAP/ Endo180 is a major pathway of extracellular matrix turnover during malignancy. J Cell Biol 169:977985. 28. Mollenhauer J, von der Mark K. 1983. Isolation and characterization of a collagen-binding glycoprotein from chondrocyte membranes. EMBO J 2:4550. 29. Kirsch T, Harrison G, Golub EE, Nah HD. 2000. The roles of annexins and types II and X collagen in matrix vesicle-mediated mineralization of growth plate cartilage. J Biol Chem 275:3557735583. 30. Pfaffle M, Ruggiero F, Hofmann H, Fernandez MP, Selmin O, et al. 1988. Biosynthesis, secretion and extracellular localization of anchorin CII, a collagen-binding protein of the calpactin family. EMBO J 7:23352342. 31. Ponta H, Sherman L, Herrlich PA. 2003. CD44: from adhesion molecules to signalling regulators. Nat Rev Mol Cell Biol 4:3345. 32. Jalkanen S, Jalkanen M. 1992. Lymphocyte CD44 binds the COOHterminal heparin-binding domain of fibronectin. J Cell Biol 116:817 825. 33. Elenius K, Salmivirta M, Inki P, Mali M, Jalkanen M. 1990. Binding of human syndecan to extracellular matrix proteins. J Biol Chem 265: 1783717843. 34. Liu W, Litwack ED, Stanley MJ, Langford JK, Lander AD, et al. 1998. Heparan sulfate proteoglycans as adhesive and anti-invasive molecules. Syndecans and glypican have distinct functions. J Biol Chem 273: 2282522832. 35. Knight CG, Morton LF, Peachey AR, Tuckwell DS, Farndale RW, et al. 2000. The collagen-binding A-domains of integrins a1b1 and a2b1 recognize the same specific amino acid sequence, GFOGER, in native (triple-helical) collagens. J Biol Chem 275:3540. 36. Knight CG, Morton LF, Onley DJ, Peachey AR, Messent AJ, et al. 1998. Identification in collagen type I of an integrin a2b1-binding site containing an essential GER sequence. J Biol Chem 273:33287 33294. pyla J, Puranen JS, Knight CG, Tiger CF, et al. 2003. 37. Zhang WM, Ka a2b1 integrin recognizes the GFOGER sequence in interstitial collagens. J Biol Chem 278:72707277. 38. Emsley J, Knight CG, Farndale RW, Barnes MJ, Liddington RC. 2000. Structural basis of collagen recognition by integrin alpha2beta1. Cell 101:4756. 39. Raynal N, Hamaia SW, Siljander PR, Maddox B, Peachey AR, et al. 2006. Use of synthetic peptides to locate novel integrin a2b1-binding motifs in human collagen III. J Biol Chem 281:38213831. 40. Eble JA, Golbik R, Mann K, Kuhn K. 1993. The alpha 1 beta 1 integrin recognition site of the basement membrane collagen molecule [a1(IV)]2 a2(IV). EMBO J 12:47954802. 41. Golbik R, Eble JA, Ries A, Kuhn K. 2000. The spatial orientation of the essential amino acid residues arginine and aspartate within the a1b1 integrin recognition site of collagen IV has been resolved using fluorescence resonance energy transfer. J Mol Biol 297:501509. 42. Sacca B, Fiori S, Moroder L. 2003. Studies of the local conformational properties of the cell-adhesion domain of collagen type IV in synthetic heterotrimeric peptides. Biochemistry 42:34293436. 43. Perret S, Eble JA, Siljander PR, Merle C, Farndale RW, et al. 2003. Prolyl hydroxylation of collagen type I is required for efficient binding to integrin a1b1 and platelet glycoprotein VI but not to a2b1. J Biol Chem 278: 2987329879. pyla J, Ja a linoja J, Tulla M, Ylo stalo J, Nissinen L, et al. 2004. The 44. Ka fibril associated collagen IX provides a novel mechanism for cell adhesion to cartilaginous matrix. J Biol Chem 279:5167751687. 45. Eble JA, Kassner A, Niland S, Morgelin M, Grifka J, et al. 2006. Collagen XVI harbors an integrin alpha1 beta1 recognition site in its C-terminal domains. J Biol Chem 281:2574525756.

46. Leitinger B, Steplewski A, Fertala A. 2004. The D2 period of collagen II contains a specific binding site for the human discoidin domain receptor, D DR2. J Mol Biol 344:9931003. 47. Kehrel B, Wierwille S, Clemetson KJ, Anders O, Steiner M, et al. 1998. Glycoprotein VI is a major collagen receptor for platelet activation: it recognizes the platelet-activating quaternary structure of collagen, whereas CD36, glycoprotein IIb/IIIa, and von Willebrand factor do not. Blood 91:491499. 48. Smethurst PA, Onley DJ, Jarvis GE, OConnor MN, Knight CG, et al. 2007. Structural basis for the platelet-collagen interaction: the smallest motif within collagen that recognizes and activates platelet Glycoprotein VI contains two glycine-proline-hydroxyproline triplets. J Biol Chem 282: 12961304. 49. Gullberg D, Gehlsen KR, Turner DC, Ahlen K, Zijenah LS, et al. 1992. Analysis of a1b1, a2b1 and a3b1 integrins in cell-collagen interactions: identification of conformation dependent a1b1 binding sites in collagen type I. EMBO J 11:38653873. pyla J, Jokinen J, et al. 2001. The 50. Nykvist P, Tasanen K, Viitasalo T, Ka cell adhesion domain of type XVII collagen promotes integrin-mediated cell spreading by a novel mechanism. J Biol Chem 276:3867338679. 51. Chen M, OToole EA, Li YY, Woodley DT. 1999. Alpha 2 beta 1 integrin mediates dermal fibroblast attachment to type VII collagen via a 158amino-acid segment of the NC1 domain. Exp Cell Res 249:231239. 52. Rehn M, Veikkola T, Kukk-Valdre E, Nakamura H, Ilmonen M, et al. 2001. Interaction of endostatin with integrins implicated in angiogenesis. Proc Natl Acad Sci USA 98:10241029. 53. Wickstrom SA, Alitalo K, Keski-Oja J. 2002. Endostatin associates with integrin alpha5beta1 and caveolin-1, and activates Src via a tyrosyl phosphatase-dependent pathway in human endothelial cells. Cancer Res 62:55805589. 54. Sudhakar A, Sugimoto H, Yang C, Lively J, Zeisberg M, et al. 2003. Human tumstatin and human endostatin exhibit distinct antiangiogenic activities mediated by avb3 and a5b1 integrins. Proc Natl Acad Sci USA 100:47664771. 55. Nyberg P, Xie L, Kalluri R. 2005. Endogenous inhibitors of angiogenesis. Cancer Res 65:39673979. 56. Sudhakar A, Nyberg P, Keshamouni VG, Mannam AP, Li J, et al. 2005. Human alpha1 type IV collagen NC1 domain exhibits distinct antiangiogenic activity mediated by a1b1 integrin. J Clin Invest 115: 28012810. 57. Magnon C, Galaup A, Mullan B, Rouffiac V, Bouquet C, et al. 2005. Canstatin acts on endothelial and tumor cells via mitochondrial damage initiated through interaction with alphavbeta3 and alphavbeta5 integrins. Cancer Res 65:43534361. 58. Borza CM, Pozzi A, Borza DB, Pedchenko V, Hellmark T, et al. 2006. Integrin a3b1, a novel receptor for a3(IV) noncollagenous domain and a trans-dominant Inhibitor for integrin avb3. J Biol Chem 281:20932 20939. 59. Tuckwell DS, Reid KB, Barnes MJ, Humphries MJ. 1996. The A-domain of integrin a2 binds specifically to a range of collagens but is not a general receptor for the collagenous motif. Eur J Biochem 241:732 739. 60. Brown JC, Mann K, Wiedemann H, Timpl R. 1993. Structure and binding properties of collagen type XIV isolated from human placenta. J Cell Biol 120:557567. 61. Klein G, Kibler C, Schermutzki F, Brown J, Muller CA, et al. 1998. Cell binding properties of collagen type XIV for human hematopoietic cells. Matrix Biol 16:307317. 62. Kassner A, Hansen U, Miosge N, Reinhardt DP, Aigner T, et al. 2003. Discrete integration of collagen XVI into tissue-specific collagen fibrils or beaded microfibrils. Matrix Biol 22:131143. 63. Kern A, Eble J, Golbik R, Kuhn K. 1993. Interaction of type IV collagen with the isolated integrins a1b1 and a2b1. Eur J Biochem 215:151 159. pyla J, Ivaska J, Riikonen R, Nykvist P, Pentika inen O, et al. 2000. 64. Ka Integrin a2I domain recognizes type I and type IV collagens by different mechanisms. J Biol Chem 275:33483354. inen O, Viitasalo T, Ka pyla J, Impola U, et al. 2001. 65. Tulla M, Pentika Selective binding of collagen subtypes by integrin a1I, a2I, and a10I domains. J Biol Chem 276:4820648212.

BioEssays 29.10

1009

Review articles

pyla J, White DJ, et al. 2004 Integrin66. Jokinen J, Dadu E, Nykvist P, Ka mediated cell adhesion to type I collagen fibrils. J Biol Chem 279: 3195631963. pyla J, Pihlajaniemi T, et al. 2000. Distinct 67. Nykvist P, Tu H, Ivaska J, Ka recognition of collagen subtypes by a1b1 and a2b1 integrins a1b1 mediates cell adhesion to type XIII collagen. J Biol Chem 275:8255 8261. 68. Saelman EU, Nieuwenhuis HK, Hese KM, de Groot PG, Heijnen HF, et al. 1994. Platelet adhesion to collagen types I through VIII under conditions of stasis and flow is mediated by GPIa/IIa (a2b1-integrin). Blood 83:12441250. 69. Siljander PR, Hamaia S, Peachey AR, Slatter DA, Smethurst PA, et al. 2004. Integrin activation state determines selectivity for novel recognition sites in fibrillar collagens. J Biol Chem 279:4776347772. 70. Velling T, Risteli J, Wennerberg K, Mosher DF, Johansson S. 2002. Polymerization of type I and III collagens is dependent on fibronectin and enhanced by integrins a11b1 and a2b1. J Biol Chem 277:37377 37381. 71. Mihai C, Iscru DF, Druhan LJ, Elton TS, Agarwal G. 2006. Discoidin domain receptor 2 inhibits fibrillogenesis of collagen type 1. J Mol Biol 361:864876. 72. Marini JC, Forlino A, Cabral WA, Barnes AM, San Antonio JD, et al. 2007. Consortium for osteogenesis imperfecta mutations in the helical domain of type I collagen: regions rich in lethal mutations align with collagen binding sites for integrins and proteoglycans. Hum Mutat 28:209221. 73. Xu Y, Gurusiddappa S, Rich RL, Owens RT, Keene DR, et al. 2000. Multiple binding sites in collagen type I for the integrins a1b1 and a2b1. J Biol Chem 275:3898138989. 74. Gardner H, Kreidberg J, Koteliansky V, Jaenisch R. 1996. Deletion of integrin alpha 1 by homologous recombination permits normal murine development but gives rise to a specific deficit in cell adhesion. Dev Biol 175:301313. 75. Ekholm E, Hankenson KD, Uusitalo H, Hiltunen A, Gardner H, et al. 2002. Diminished callus size and cartilage synthesis in a1b1 integrin deficient mice during bone fracture healing. Am J Pathol 160:1779 1785. 76. Zemmyo M, Meharra EJ, Kuhn K, Creighton-Achermann L, Lotz M. 2003. Accelerated, aging-dependent development of osteoarthritis in alpha1 integrin-deficient mice. Arthritis Rheum 48:28732880. 77. Pozzi A, Wary KK, Giancotti FG, Gardner HA. 1998. Integrin alpha1beta1 mediates a unique collagen-dependent proliferation pathway in vivo. J Cell Biol 142:587594. 78. Gardner H, Broberg A, Pozzi A, Laato M, Heino J. 1999. Absence of integrin a1b1 in the mouse causes loss of feedback regulation of collagen synthesis in normal and wounded dermis. J Cell Sci 112:263 272. 79. Chen X, Moeckel G, Morrow JD, Cosgrove D, Harris RC, et al. 2004. Lack of integrin alpha1beta1 leads to severe glomerulosclerosis after glomerular injury. Am J Pathol 165:617630. 80. Pozzi A, Moberg PE, Miles LA, Wagner S, Soloway P, et al. 2000. Elevated matrix metalloprotease and angiostatin levels in integrin a1 knockout mice cause reduced tumor vascularization. Proc Natl Acad Sci USA 97:22022207. 81. Senger DR, Perruzzi CA, Streit M, Koteliansky VE, de Fougerolles AR, et al. The a1b1 and a2b1 integrins provide critical support for vascular endothelial growth factor signaling, endothelial cell migration, and tumor angiogenesis. Am J Pathol 160:195204. 82. Rubio MA, Sotillos M, Jochems G, Alvarez V, Corbi AL. 1995. Monocyte activation: rapid induction of a1/b1 (VLA-1) integrin expression by lipopolysaccharide and interferon-gamma. Eur J Immunol 25:2701 2705.

83. Fiorucci S, Mencarelli A, Palazzetti B, Sprague AG, Distrutti E, et al. 2002. Importance of innate immunity and collagen binding integrin a1b1 in TNBS-induced colitis. Immunity 17:769780. 84. Ray SJ, Franki SN, Pierce RH, Dimitrova S, Koteliansky V, et al. 2004. The collagen binding alpha1beta1 integrin VLA-1 regulates CD8 T cellmediated immune protection against heterologous influenza infection. Immunity 20:167179. 85. Holtkotter O, Nieswandt B, Smyth N, Muller W, Hafner M, et al. 2002. Integrin alpha 2-deficient mice develop normally, are fertile, but display partially defective platelet interaction with collagen. J Biol Chem 277:1078910794. 86. Chen J, Diacovo TG, Grenache DG, Santoro SA, Zutter MM. 2002. The a(2) integrin subunit-deficient mouse: a multifaceted phenotype including defects of branching morphogenesis and hemostasis. Am J Pathol 161:337344. 87. He L, Pappan LK, Grenache DG, Li Z, Tollefsen DM, et al. 2003. The contributions of the alpha 2 beta 1 integrin to vascular thrombosis in vivo. Blood 102:36523657. 88. Edelson BT, Li Z, Pappan LK, Zutter MM. 2004. Mast cell-mediated inflammatory responses require the a2b1 integrin. Blood 103:2214 2220. 89. Grenache DG, Zhang Z, Wells LE, Santoro SA, Davidson JM, et al. 2007. Wound healing in the a2b1 integrin-deficient mouse: altered keratinocyte biology and dysregulated matrix metalloproteinase expression. J Invest Dermatol 127:455466. 90. Zweers MC, Davidson JM, Pozzi A, Hallinger R, Janz K, et al. 2007. Integrin a2b1 is required for regulation of murine wound angiogenesis but is dispensable for reepithelialization. J Invest Dermatol 127:467 478. 91. Larjava H, Salo T, Haapasalmi K, Kramer R, Heino J. 1993. Expression of integrins and basement membrane components by wound keratinocytes. J Clin Invest 92:14251435. 92. Bengtsson T, Aszodi A, Nicolae C, Hunziker EB, Lundgren-Akerlund E, et al. 2005. Loss of a10b1 integrin expression leads to moderate dysfunction of growth plate chondrocytes. J Cell Sci 118:929 936. 93. Popova SN, Barczyk M, Tiger CF, Beertsen W, Zigrino P, et al. 2007. alpha11beta1 integrin-dependent regulation of periodontal ligament function in the erupting mouse incisor. Mol Cell Biol 27:43064316. 94. Vogel WF, Aszodi A, Alves F, Pawson T. 2001. Discoidin domain receptor 1 tyrosine kinase has an essential role in mammary gland development. Mol Cell Biol 21:29062917. 95. Gross O, Beirowski B, Harvey SJ, McFadden C, Chen D, et al. 2004. DDR1-deficient mice show localized subepithelial GBM thickening with focal loss of slit diaphragms and proteinuria. Kidney Int 66:102 111. 96. Labrador JP, Azcoitia V, Tuckermann J, Lin C, Olaso E, et al. 2001. The collagen receptor DDR2 regulates proliferation and its elimination leads to dwarfism. EMBO Rep 2:446452. pez-Ot ha ri VM. 1999. Induction of n C, Ka 97. Ravanti L, Heino J, Lo collagenase-3 (MMP-13) expression in human skin fibroblasts by three-dimensional collagen is mediated by p38 mitogen-activated protein kinase. J Biol Chem 274:24462455. 98. Parks WC. 2007. What is the alpha2beta1 integrin doing in the epidermis? J Invest Dermatol 127:264266. 99. Lee W, Sodek J, McCulloch CA. 1996. Role of integrins in regulation of collagen phagocytosis by human fibroblasts. J Cell Physiol 168:695 704. ki V, Kankaanpa a P, Ivaska J, Hyypia T, et al. 2004. 100. Upla P, Marjoma Clustering induces a lateral redistribution of a2b1 integrin from membrane rafts to caveolae and PKC-dependent internalization. Mol Biol Cell 15:625636.

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BioEssays 29.10