Guanosine 5'-O-(3-thiotriphosphate) (GTPγS) stimulation of GLUT4 translocation is tyrosine kinase-dependent

Jeffrey S. Elmendorf, Dong Chen, Jeffrey E. Pessin

Research output: Contribution to journalArticle

65 Scopus citations

Abstract

Guanosine 5'-O-(3-thiotriphosphate) (GTPγS) treatment of permeabilized adipocytes results in GLUT4 translocation similar to that elicited by insulin treatment. However, although the selective phosphatidylinositol 3-kinase inhibitor, wortmannin, completely prevented insulin-stimulated GLUT4 translocation, it was without effect on GTPγS-stimulated GLUT4 translocation. In addition, insulin was an effective stimulant, whereas GTPγS was a very weak activator of the downstream Akt serine/threonine kinase. Consistent with an Akt-independent mechanism, guanosine 5'-O-2- (thio) diphosphate inhibited insulin-stimulated GLUT4 translocation without any effect on the Akt kinase. Surprisingly, two functionally distinct tyrosine kinase inhibitors, genistein and herbimycin A, as well as microinjection of a monoclonal phosphotyrosine specific antibody, inhibited both GTPγS- and insulin-stimulated GLUT4 translocation. Phosphotyrosine immunoblotting and specific immunoprecipitation demonstrated that GTPγS did not elicit tyrosine phosphorylation of insulin receptor or insulin receptor substrate-1. In contrast to insulin, proteins in the 120-130-kDa and 55-75- kDa range were tyrosine-phosphorylated following GTPγS stimulation. Several of these proteins were identified and include protein-tyrosine kinase 2 (also known as CAKβ, RAFTK, and CADTK), pp125 focal adhesion tyrosine kinase, pp130 Crk-associated substrate, paxillin, and Cbl. These data demonstrate that the GTPγS-stimulated GLUT4 translocation utilizes a novel tyrosine kinase pathway that is independent of both the phosphatidylinositol 3-kinase and the Akt kinase.

Original languageEnglish (US)
Pages (from-to)13289-13296
Number of pages8
JournalJournal of Biological Chemistry
Volume273
Issue number21
DOIs
StatePublished - May 22 1998
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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