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Purification and characterization of G proteins from human brain: modification of GTPase activity upon phosphorylation

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Summary

Three G proteins from human brain membranes were purified to near homogeneity by conventional techniques including preparative electrophoresis. These G proteins were characterized by their ability to bind GTP, GDP and GTP analogs. Two of these proteins have molecular weights of 50,000 (G50) and 36,000 (G36), as determined on SDS-gels. G36 was ADP-ribosylated by pertussis toxin. Thus, G50 could represent a Gsα subunit, whereas G36 could be Giα or Goα. G50 was phosphorylated by cAMP dependent protein kinase and protein kinase C. G36 was phosphorylated by a protein kinase independent of calcium and phospholipid, a proteolytic product of protein kinase C, analogous to protein kinase M. Phosphorylation of G36 by this protein kinase induced a dramatic decrease in its GTPase activity. The third G protein, of molecular weight 22,000 probably belongs to the group of monomeric G proteins possessing functional similarities withras gene products. The regulation of G proteins involving calcium-dependent and independent pathways is delineated.

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Authors and Affiliations

  1. Centre de Recherche Delalande, 10 rue des Carrières, 92500, Rueil-Malmaison, France

    Claude Sauvage & Jean-François Rumigny

  2. Centre de Neurochimie du CNRS, 5 rue Blaise Pascal, 67084, Strasbourg Cedex, France

    Claude Sauvage & Michel Maitre

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  1. Claude Sauvage
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  2. Jean-François Rumigny
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  3. Michel Maitre
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Sauvage, C., Rumigny, JF. & Maitre, M. Purification and characterization of G proteins from human brain: modification of GTPase activity upon phosphorylation. Mol Cell Biochem 107, 65–77 (1991). https://doi.org/10.1007/BF02424577

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  • DOI: https://doi.org/10.1007/BF02424577

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