title: Kinetic regulation of beta 3 integrin tyrosine phosphorylation.
abstract: Tyrosine phosphorylation of beta(3) integrins is a permissive stage in the activation of alpha(IIb)beta(3) and alpha(v)beta(3) in platelets and leukocytes, respectively. In this study we demonstrated direct phosphorylation of beta(3) integrins as a result of interaction with soluble monomeric ligand, and we characterized the differential kinetics of beta(3) phosphorylation as a consequence of alpha subunit pairing. We found that beta(3) phosphorylation is initiated by RGD peptide binding in a dose-dependent and saturable fashion with alpha(IIb)beta(3) becoming phosphorylated and dephosphorylated more rapidly than alpha(v)beta(3). Site mapping of phosphate incorporation reveals significant phosphorylation at Tyr-747 in both beta(3) integrin species with incorporation at Tyr-759 found at significant levels only in alpha(IIb)beta(3). Mutation of cytoplasmic beta(3) tyrosine residues in a transfection model prevents cell adhesion via these integrins. These data demonstrate that recognition of ligand is sufficient to induce beta(3) tyrosine phosphorylation and suggests that this event is regulated by the alpha subunit pairing of beta(3).
mesh_terms: D000818:Animals; D001665:Binding Sites; D002448:Cell Adhesion; D003593:Cytoplasm / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005434:Flow Cytometry; D005786:Gene Expression Regulation; D006801:Humans; D039302:Integrin alphaVbeta3 / Q000378:metabolism; D039661:Integrin beta3 / Q000737:chemistry* / Q000378:metabolism*; D020014:K562 Cells; D007700:Kinetics; D008024:Ligands; D051379:Mice; D009154:Mutation; D009842:Oligopeptides / Q000494:pharmacology; D010455:Peptides / Q000737:chemistry; D010766:Phosphorylation; D011485:Protein Binding; D013997:Time Factors; D014162:Transfection; D014443:Tyrosine / Q000378:metabolism*
chemical_list: D039302:Integrin alphaVbeta3; D039661:Integrin beta3; D008024:Ligands; D009842:Oligopeptides; D010455:Peptides; D014443:Tyrosine; C047981:arginyl-glycyl-aspartic acid