title: Cooperative role of the membrane-proximal and -distal residues of the integrin beta3 cytoplasmic domain in regulation of talin-mediated alpha IIb beta3 activation.
abstract: Integrin cytoplasmic tails regulate integrin activation that is required for high affinity binding with ligands. The interaction of the integrin beta subunit tail with a cytoplasmic protein, talin, largely contributes to integrin activation. Here we report the cooperative interaction of the beta3 membrane-proximal and -distal residues in regulation of talin-mediated alpha IIb beta3 activation. Because a chimeric integrin, alpha IIb beta3/beta1, in which the beta3 tail was replaced with the beta1 tail was constitutively active, we searched for the residues responsible for integrin activation among the residues that differed between the beta3 and beta1 tails. Single amino acid substitutions of Ile-719 and Glu-749 in the beta3 membrane-proximal and -distal regions, respectively, with the corresponding beta1 residues or alanine rendered alphaIIbbeta3 constitutively active. The I719M/E749S double mutant had the same ligand binding activity as alpha IIb beta3/beta1. These beta3 mutations also induced alphaVbeta3 activation. Conversely, substitution of Met-719 or Ser-749 in the beta1 tail with the corresponding beta3 tail residue (M719I or S749E) inhibited alpha IIb beta3/beta1 activation, and the M719I/S749E double mutant inhibited ligand binding to a level comparable with that of the wild-type alpha IIb beta3. Knock down of talin by short hairpin RNA inhibited the I719M- and E749S-induced alpha IIb beta3 activation. These results suggest that the beta3 membrane-proximal and -distal residues cooperatively regulate talin-mediated alpha IIb beta3 activation.
authors.affiliation: Division of Blood Transfusion, Ehime University Graduate School of Medicine, Ehime 791-0295, Japan. takahato@m.ehime-u.ac.jp
authors.forename: Takaaki
authors.initials: T
authors.lastname: Hato
authors.forename: Jun
authors.initials: J
authors.lastname: Yamanouchi
authors.forename: Tatsushiro
authors.initials: T
authors.lastname: Tamura
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Yakushijin
authors.forename: Ikuya
authors.initials: I
authors.lastname: Sakai
authors.forename: Masaki
authors.initials: M
authors.lastname: Yasukawa
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D006801:Humans; D039661:Integrin beta3 / Q000235:genetics / Q000378:metabolism*; D050506:Mutant Chimeric Proteins / Q000235:genetics / Q000378:metabolism; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding / Q000502:physiology; D017434:Protein Structure, Tertiary / Q000502:physiology; D016608:Talin / Q000235:genetics / Q000378:metabolism*
chemical_list: D039661:Integrin beta3; D050506:Mutant Chimeric Proteins; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D016608:Talin