Arginine-glycine-aspartic acid binding leading to molecular stabilization between integrin alpha v beta 3 and its ligand.
Cell adhesion is characterized by an integrin-mediated ligand binding event followed by reorganization of the actin-cytoskeleton leading to cell spreading and/or migration. In this report we examine the role of integrin alpha v beta 3 in mediating cell attachment to vitronectin or a RGD-containing peptide in the presence of cytochalasin B to prevent actin polymerization. Under these conditions cell attachment to a RGD-containing peptide can be dissociated by excess soluble ligand whereas cells attached to vitronectin cannot. These results suggest that alpha v beta 3-mediated cell attachment to vitronectin results in a highly stabilized interaction that is independent of the actin-cytoskeleton. To investigate the molecular nature of this interaction alpha v beta 3 was purified to homogeneity, and its binding properties toward various ligands were measured in a solid-phase receptor assay. The data indicate that alpha v beta 3 binds to vitronectin or fibronectin in a nondissociable manner whereas a RGD-containing peptide derived from vitronectin binds specifically but is completely dissociable with a Kd of 9.4 x 10(-7) M. Moreover, chemical modification of alpha v beta 3 with limited glutaraldehyde treatment allowed vitronectin to bind in a RGD-dependent and dissociable manner, suggesting that receptor conformational changes or specific amino acid residues proximal to the ligand binding site(s) are involved in the stabilization event. Thus, in the absence of cytoskeletal proteins or other cellular components, integrin alpha v beta 3-ligand binding involves recognition of the RGD sequence leading to a highly stabilized protein-protein association.
Orlando
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RA
Department of Immunology, Scripps Research Institute, La Jolla, California 92037.
Cheresh
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DA
D000595:Amino Acid Sequence; D002448:Cell Adhesion / Q000187:drug effects; D003571:Cytochalasin B / Q000494:pharmacology; D005976:Glutaral / Q000378:metabolism; D006023:Glycoproteins / Q000378:metabolism; D006801:Humans; D016023:Integrins / Q000378:metabolism*; D007700:Kinetics; D008024:Ligands; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000378:metabolism*; D014407:Tumor Cells, Cultured / Q000187:drug effects; D019096:Vitronectin
D006023:Glycoproteins; D016023:Integrins; D008024:Ligands; D009842:Oligopeptides; D019096:Vitronectin; D003571:Cytochalasin B; C047981:arginyl-glycyl-aspartic acid; D005976:Glutaral