Photoaffinity labeling of two rat liver plasma membrane proteins with [32P]γ-azidoanilido GTP in response to vasopressin: Immunologic identification as α subunits of the Gq class of G proteins

Ronald L. Wange, Alan V. Smrcka, Paul C. Sternweis, John H. Exton

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Abstract

Two proteins have been identified in rat liver plasma membranes that bind a photoreactive GTP analogue, [32P]γ-azidoanilido GTP, in response to incubation with the Ca2+-mobilizing agonist, vasopressin. The labeled proteins possess apparent molecular masses of 42 and 43 kDa. Their labeling requires Mg2+ and can be inhibited by GTP, its analogues, and GDP but not by other nucleotides. Vasopressin-stimulated labeling is attenuated by a V1 receptor-selective antagonist. The concentration of vasopressin required to stimulate labeling is in the same range (EC50 = 4 nM) as that required for activation of GTPase and phosphoinositide-specific phospholipase C activities in liver plasma membranes. Immunodetection and immunoprecipitation of the [32P]γ-azidoanilido GTP-labeled 42- and 43-kDa proteins with antisera raised against peptide sequences in αq indicate that these proteins are members of the recently described Gq class of G proteins.

Original languageEnglish (US)
Pages (from-to)11409-11412
Number of pages4
JournalJournal of Biological Chemistry
Volume266
Issue number18
Publication statusPublished - 1991

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ASJC Scopus subject areas

  • Biochemistry

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