title: Interactions of platelet integrin alphaIIb and beta3 transmembrane domains in mammalian cell membranes and their role in integrin activation.
abstract: Clustering and occupancy of platelet integrin alpha(IIb)beta(3) (GPIIb-IIIa) generate biologically important signals: conversely, intracellular signals increase the integrins' affinity, leading to integrin activation; both forms of integrin signaling play important roles in hemostasis and thrombosis. Indirect evidence implicates interactions between integrin alpha and beta transmembrane domains (TMDs) and cytoplasmic domains in integrin signaling; however, efforts to directly identify these associations have met with varying and controversial results. In this study, we develop mini-integrin affinity capture and use it in combination with nuclear magnetic resonance spectroscopy to show preferential heterodimeric association of integrin alpha(IIb)beta(3) TMD tails via specific TMD interactions in mammalian cell membranes in lipid bicelles. Furthermore, charge reversal mutations at alpha(IIb)(R995)beta(3)(D723) confirm a proposed salt bridge and show that it stabilizes the TMD-tail association; talin binding to the beta(3) tail, which activates the integrin, disrupts this association. These studies establish the preferential heterodimeric interactions of integrin alpha(IIb)beta(3) TMD tails in mammalian cell membranes and document their role in integrin signaling.
authors.affiliation: Department of Medicine, University of California San Diego, La Jolla, CA 92093, USA.
authors.forename: Chungho
authors.initials: C
authors.lastname: Kim
authors.forename: Tong-Lay
authors.initials: TL
authors.lastname: Lau
authors.forename: Tobias S
authors.initials: TS
authors.lastname: Ulmer
authors.forename: Mark H
authors.initials: MH
authors.lastname: Ginsberg
mesh_terms: D000818:Animals; D001792:Blood Platelets / Q000378:metabolism*; D016466:CHO Cells; D002462:Cell Membrane / Q000378:metabolism*; D006224:Cricetinae; D003412:Cricetulus; D003593:Cytoplasm / Q000378:metabolism; D019281:Dimerization; D005434:Flow Cytometry; D009682:Magnetic Resonance Spectroscopy; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000737:chemistry* / Q000378:metabolism*; D040201:Platelet Membrane Glycoprotein IIb / Q000737:chemistry* / Q000378:metabolism*; D011485:Protein Binding
chemical_list: D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D040201:Platelet Membrane Glycoprotein IIb