title: Structural analysis of the alpha(2) integrin I domain/procollagenase-1 (matrix metalloproteinase-1) interaction.
abstract: Previous studies have established that ligation of keratinocyte alpha(2)beta(1) integrin by type I collagen induces expression of matrix metalloproteinase-1 (MMP-1) and that MMP-1 activity is required for the alpha(2)beta(1) integrin-dependent migration of primary keratinocytes across collagenous matrices. We now present evidence that MMP-1 binds the alpha(2)beta(1) integrin via the I domain of the alpha(2) integrin subunit. Using an enzyme-linked immunosorbent assay with purified human MMP-1 and recombinant alpha(2) integrin I domain, we showed that the alpha(2) integrin I domain specifically bound in a divalent cation-dependent manner to both the pro and active forms of MMP-1, but not to MMP-3 or MMP-13. Although both the I domain and MMP-1 bind divalent cations, MMP-1 bound, in a divalent cation-dependent manner, to alpha(2) integrin I domains containing metal ion-dependent adhesion sites motif mutations that prevent divalent cation binding to the I domain, demonstrating that the metal ion dependence is a function of MMP-1. Using a series of MMP-1-MMP-3 and MMP-1-MMP-13 chimeras, we determined that both the linker domain and the hemopexin-like domain of MMP-1 were required for optimal binding to the I domain. The alpha(2) integrin/MMP-1 interaction described here extends an emerging paradigm in matrix biology involving anchoring of proteinases to the cell surface to regulate their biological activities.
authors.affiliation: Departments of Pathology and Immunology, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
authors.forename: T P
authors.initials: TP
authors.lastname: Stricker
authors.forename: J A
authors.initials: JA
authors.lastname: Dumin
authors.forename: S K
authors.initials: SK
authors.lastname: Dickeson
authors.forename: L
authors.initials: L
authors.lastname: Chung
authors.forename: H
authors.initials: H
authors.lastname: Nagase
authors.forename: W C
authors.initials: WC
authors.lastname: Parks
authors.forename: S A
authors.initials: SA
authors.lastname: Santoro
mesh_terms: D015703:Antigens, CD / Q000737:chemistry* / Q000378:metabolism*; D001665:Binding Sites; D002413:Cations, Divalent / Q000494:pharmacology; D002465:Cell Movement; D003001:Cloning, Molecular; D003094:Collagen / Q000502:physiology; D017364:Collagenases / Q000737:chemistry* / Q000378:metabolism*; D004792:Enzyme Precursors / Q000737:chemistry* / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D039421:Integrin alpha2; D015603:Keratinocytes / Q000502:physiology; D007700:Kinetics; D020781:Matrix Metalloproteinase 1 / Q000737:chemistry* / Q000378:metabolism*; D053509:Matrix Metalloproteinase 13; D019278:Matrix Metalloproteinase 3 / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism
chemical_list: D015703:Antigens, CD; D002413:Cations, Divalent; D004792:Enzyme Precursors; D039421:Integrin alpha2; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D003094:Collagen; D017364:Collagenases; C507079:MMP13 protein, human; D053509:Matrix Metalloproteinase 13; D019278:Matrix Metalloproteinase 3; D020781:Matrix Metalloproteinase 1