Multiple Binding Sites in Collagen Type I for the Integrins α1β1 and α2β1

Xu, Yi; Gurusiddappa, Sivashankarappa; Rich, Rebecca L.; Owens, Rick T.; Keene, Douglas R.; Mayne, Richard; Höök, Agneta; Höök, Magnus

doi:10.1074/jbc.m007668200

Cited by 229 publications

(263 citation statements)

References 48 publications

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“…These binding sites share a GER motif, but variations in the flanking sequences may modulate their affinity for ␣ 2 ␤ 1 . However, the narrow, unimodal rupture force distribution we obtained during single-molecule measurements suggests that ␣2␤1 bound to all accessible binding sites with similar affinity, consistent with previous observations (Xu et al, 2000).…”

Section: Discussionsupporting

confidence: 80%

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Revealing Early Steps of α₂β₁Integrin-mediated Adhesion to Collagen Type I by Using Single-Cell Force Spectroscopy

Taubenberger

Cisneros

Friedrichs

et al. 2007

MBoC

193

197

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We have characterized early steps of ␣ 2 ␤ 1 integrin-mediated cell adhesion to a collagen type I matrix by using single-cell force spectroscopy. In agreement with the role of ␣ 2 ␤ 1 as a collagen type I receptor, ␣ 2 ␤ 1 -expressing Chinese hamster ovary (CHO)-A2 cells spread rapidly on the matrix, whereas ␣ 2 ␤ 1 -negative CHO wild-type cells adhered poorly. Probing CHO-A2 cell detachment forces over a contact time range of 600 s revealed a nonlinear adhesion response. During the first 60 s, cell adhesion increased slowly, and forces associated with the smallest rupture events were consistent with the breakage of individual integrin-collagen bonds. Above 60 s, a fraction of cells rapidly switched into an activated adhesion state marked by up to 10-fold increased detachment forces. Elevated overall cell adhesion coincided with a rise of the smallest rupture forces above the value required to break a single-integrin-collagen bond, suggesting a change from single to cooperative receptor binding. Transition into the activated adhesion mode and the increase of the smallest rupture forces were both blocked by inhibitors of actomyosin contractility. We therefore propose a two-step mechanism for the establishment of ␣ 2 ␤ 1 -mediated adhesion as weak initial, single-integrin-mediated binding events are superseded by strong adhesive interactions involving receptor cooperativity and actomyosin contractility. INTRODUCTIONIntegrins are a family of ␣/␤-heterodimeric receptors involved in cell-cell adhesion and cell attachment to the extracellular matrix (Hynes, 1992). In adherent cells, integrins are often arranged into highly organized structures, such as focal complexes, focal adhesions, and fibrillar adhesions . In these mature adhesive contacts, integrins are linked to the actin cytoskeleton via their intracellular domains and a multitude of adapter proteins, such as vinculin, talin, and ␣-actinin (Zamir and Geiger, 2001). Whereas the molecular composition of integrin complexes and signaling pathways controlling their macroscopic assembly or disassembly have been analyzed in detail, less is known about the early molecular events leading to the formation of integrin-based adhesion.Generally, integrin-mediated adhesion is thought to be initiated on the cell membrane by the engagement of individual integrin dimers with their respective ligand (Lotz et al., 1989;Gallant and Garcia, 2006). The number of receptorligand pairs may then grow by increasing the cell-substrate contact area and by receptor diffusion within that zone. Subsequent adhesion strengthening occurs through integrin clustering and linkage to the cytoskeleton. In addition, the ligand affinity of the integrins may increase as a result of intracellular regulatory pathways (Hughes and Pfaff, 1998). At a later stage, the assembly of higher-order integrincytoskeletal complexes requires actomyosin-driven contractility under the control of the small guanosine triphosphatase (GTPase) RhoA (Chrzanowska-Wodnicka and Burridge, 1996) and its downstream target Rho-assoc...

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Section: Discussionsupporting

confidence: 80%

“…There are several high-affinity bindings sites for ␣ 2 ␤ 1 on fibrillar collagen type I Xu et al, 2000). These binding sites share a GER motif, but variations in the flanking sequences may modulate their affinity for ␣ 2 ␤ 1 .…”

Section: Discussionmentioning

confidence: 99%

Revealing Early Steps of α₂β₁Integrin-mediated Adhesion to Collagen Type I by Using Single-Cell Force Spectroscopy

Taubenberger

Cisneros

Friedrichs

et al. 2007

MBoC

193

197

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“…4). MLBR 2 extends from helical position 682-830, and includes sites important for collagen self-assembly [Prockop and Fertala, 1998], for cleavage by matrix metalloproteinases (MMPs) 1, 2, and 13 [Lauer-Fields et al, 2000], as well as for binding by α1β1/α2β1 integrins [Xu et al, 2000], fibronectin [Dzamba et al, 1993;Kleinman and McGoodwin, 1976;Kleinman et al, 1978], cartilage oligomeric matrix protein (COMP) [Rosenberg et al, 1998], phosphophoryn [Dahl et al, 1998], and possibly the discoidin domain (DDR2) receptor [Schlessinger, 1997;Shrivastava et al, 1997;Vogel et al, 1997]. MLBR 3 extends from helical residue 920 to the end of the helical region.…”

Section: Genotype-phenotype Correlationsmentioning

confidence: 99%

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

et al. 2007

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Osteogenesis imperfecta (OI) is a generalized disorder of connective tissue characterized by fragile bones and easy susceptibility to fracture. Most cases of OI are caused by mutations in type I collagen. We have identified and assembled structural mutations in type I collagen genes (COL1A1 and COL1A2, encoding the proα1(I) and proα2(I) chains, respectively) that result in OI. Quantitative defects causing type I OI were not included. Of these 832 independent mutations, 682 result in substitution for glycine residues in the triple helical domain of the encoded protein and 150 alter splice sites. Distinct genotype-phenotype relationships emerge for each chain. Onethird of the mutations that result in glycine substitutions in α1(I) are lethal, especially when the substituting residues are charged or have a branched side chain. Substitutions in the first 200 residues are nonlethal and have variable outcome thereafter, unrelated to folding or helix stability domains. Two exclusively lethal regions (helix positions 691-823 and 910-964) align with major ligand binding regions (MLBRs), suggesting crucial interactions of collagen monomers or fibrils with integrins, matrix metalloproteinases (MMPs), fibronectin, and cartilage oligomeric matrix protein (COMP). Mutations in COL1A2 are predominantly nonlethal (80%). Lethal substitutions are located in eight regularly spaced clusters along the chain, supporting a regional model. The lethal regions align with proteoglycan binding sites along the fibril, suggesting a role in fibrilmatrix interactions. Recurrences at the same site in α2(I) are generally concordant for outcome, unlike α1(I). Splice site mutations comprise 20% of helical mutations identified in OI patients, and may lead to exon skipping, intron inclusion, or the activation of cryptic splice sites. Splice site mutations in COL1A1 are rarely lethal; they often lead to frameshifts and the mild type I phenotype. In α2(I), lethal exon skipping events are located in the carboxyl half of the chain. Our data on genotype-phenotype relationships indicate that the two collagen chains play very different roles in matrix integrity and that phenotype depends on intracellular and extracellular events.

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“…With fibroblast cellular remodeling having such critical effects on healing and cancer applications, gaining a better understanding of the fibroblastic remodeling response is important. α1β1 and α2β1 integrins are involved in binding to native collagen type I while αvβ3 is activated in response to denatured collagen type I [56,[59][60][61][62]. The α2β1 integrin binds to the GFOGER sequence in native collagen type I and does not bind to denatured collagen type I [60,62,[63][64][65].…”

Section: Discussionmentioning

confidence: 99%

“…α1β1 and α2β1 integrins are involved in binding to native collagen type I while αvβ3 is activated in response to denatured collagen type I [56,[59][60][61][62]. The α2β1 integrin binds to the GFOGER sequence in native collagen type I and does not bind to denatured collagen type I [60,62,[63][64][65]. The αvβ3 integrin binds to denatured collagen type I almost exclusively via matricryptic RGD sequences that are exposed upon denaturation and unwinding of the triple helix [59,61,[66][67][68].…”

Section: Discussionmentioning

confidence: 99%

Phagocytosis and remodeling of collagen matrices

Abraham

Dice

Lee

et al. 2007

Experimental Cell Research

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The biodegradation of collagen and the deposition of new collagen-based extracellular matrices are of central importance in tissue remodeling and function. Similarly, for collagen-based biomaterials used in tissue engineering, the degradation of collagen scaffolds with accompanying cellular infiltration and generation of new extracellular matrix is critical for integration of in vitro grown tissues in vivo. In earlier studies we observed significant impact of collagen structure on primary lung fibroblast behavior in vitro in terms of collagen uptake and matrix remodeling. Therefore, in the present work, the response of human fibroblasts (IMR-90) to the structural state of collagen was studied with respect to phagocytosis in the presence and absence of inhibitors. Protein content and transcript levels for Collagen I (Col-1), Matrix Metalloproteinase 1 (MMP-1), Matrix Metalloproteinase 2 (MMP-2), Tissue Inhibitor of Matrix Metalloproteinase 1 (TIMP-1), Tissue Inhibitor of Matrix Metalloproteinase 2 (TIMP-2), and Heat Shock Protein 70 (HSP-70) were characterized as a function of collagen matrix concentration, structure and cell culture time to assess effects on cellular collagen matrix remodeling processes. Phagocytosis of collagen was assessed quantitatively by uptake of collagen coated fluorescent beads incorporated into the pre-formed collagen matrices. Significantly higher levels of collagen phagocytosis were observed for the cells grown on the denatured collagen vs. native collagen matrices. Significant reduction in collagen phagocytosis was observed by blocking several phagocytosis pathways when the cells were grown on denatured collagen versus non-denatured collagen. Collagen phagocytosis inhibition effects were significantly greater for PDL57 IMR-90 cells versus PDL48 cells, reflecting a reduced number of collagen processing pathways available to the older cells. Transcript levels related to the deposition of new extracellular matrix proteins varied as a function of the structure of the collagen matrix presented to the cells. A four-fold increase in transcript level of Col-1 and a higher level of collagen matrix incorporation were observed for cells grown on denatured collagen versus cells grown on non-denatured collagen. The data suggest that biomaterial matrices incorporating denatured collagen may promote more active remodeling toward new extracellular matrices in comparison to cells grown on non-denatured collagen. A similar effect of cellular action toward denatured (wound-related) collagen in the remodeling of tissues in vivo may have significant impact on tissue regeneration as well as the progression of collagen-related diseases.

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Multiple Binding Sites in Collagen Type I for the Integrins α1β1 and α2β1

Cited by 229 publications

References 48 publications

Revealing Early Steps of α₂β₁Integrin-mediated Adhesion to Collagen Type I by Using Single-Cell Force Spectroscopy

Revealing Early Steps of α₂β₁Integrin-mediated Adhesion to Collagen Type I by Using Single-Cell Force Spectroscopy

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

Phagocytosis and remodeling of collagen matrices

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Multiple Binding Sites in Collagen Type I for the Integrins α1β1 and α2β1

Cited by 229 publications

References 48 publications

Revealing Early Steps of α2β1Integrin-mediated Adhesion to Collagen Type I by Using Single-Cell Force Spectroscopy

Revealing Early Steps of α2β1Integrin-mediated Adhesion to Collagen Type I by Using Single-Cell Force Spectroscopy

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

Phagocytosis and remodeling of collagen matrices

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Product

Resources

About

Revealing Early Steps of α₂β₁Integrin-mediated Adhesion to Collagen Type I by Using Single-Cell Force Spectroscopy

Revealing Early Steps of α₂β₁Integrin-mediated Adhesion to Collagen Type I by Using Single-Cell Force Spectroscopy