title: Contributions of the I and EF hand domains to the divalent cation-dependent collagen binding activity of the alpha2beta1 integrin.
abstract: The alpha2beta1 integrin binds collagen in a Mg2+-dependent manner that is inhibited by Ca2+. Like the intact integrin, purified recombinant proteins containing the alpha2 integrin I domain, either alone or with variable numbers of alpha2 integrin EF hand metal binding sites, bound collagen in a Mg2+-dependent manner, and Ca2+ did not support binding. However, unlike the intact integrin, Ca2+ did not inhibit the Mg2+-dependent binding of any of the fusion proteins to collagen. Binding to collagen was saturable and blocked by the alpha2beta1 function blocking antibody 6F1. Deletional analysis demonstrated that residues present within the amino-terminal 35 amino acids contribute to the 6F1 epitope and are required for Mg2+-dependent collagen binding. The results indicate that the I domain contains a Mg2+ binding site that is essential for collagen binding and that the I domain alone is sufficient for collagen binding. Binding is markedly enhanced in a divalent cation-dependent manner by the addition of the first EF hand motif. Mutation of the EF hand to an inactive form completely abrogated the effect. The sites necessary for Ca2+ inhibition are not present within the I domain or the adjacent region containing the three EF hand sites.
authors.affiliation: Department of Pathology, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
authors.forename: S K
authors.initials: SK
authors.lastname: Dickeson
authors.forename: J J
authors.initials: JJ
authors.lastname: Walsh
authors.forename: S A
authors.initials: SA
authors.lastname: Santoro
mesh_terms: D001665:Binding Sites; D002118:Calcium / Q000378:metabolism*; D003094:Collagen / Q000378:metabolism*; D006801:Humans; D016023:Integrins / Q000378:metabolism*; D008274:Magnesium / Q000378:metabolism*; D011485:Protein Binding; D038981:Receptors, Collagen; D011994:Recombinant Proteins / Q000378:metabolism
chemical_list: D016023:Integrins; D038981:Receptors, Collagen; D011994:Recombinant Proteins; D003094:Collagen; D008274:Magnesium; D002118:Calcium