Insulin-mediated targeting of phosphatidylinositol 3-kinase to GLUT4-containing vesicles
Program in Molecular Medicine; Department of Biochemistry and Molecular Biology
1-Phosphatidylinositol 3-Kinase; 3T3 Cells; Adipocytes; Animals; Cell Compartmentation; Enzyme Activation; Glucose Transporter Type 4; Insulin; Mice; Monosaccharide Transport Proteins; *Muscle Proteins; Organelles; Phosphoproteins; Phosphotransferases (Alcohol Group Acceptor); Rats; Receptor, Insulin; Signal Transduction
Life Sciences | Medicine and Health Sciences
Phosphatidylinositol (PI) 3-kinase is hypothesized to be a signaling element in the acute redistribution of intracellular GLUT4 glucose transporters to the plasma membrane in response to insulin. However, some receptors activate PI 3-kinase without causing GLUT4 translocation, suggesting specific cellular localization may be critical to this PI 3-kinase function. Consistent with this idea, complexes containing PI 3-kinase bound to insulin receptor substrate 1 (IRS-1) in 3T3-L1 adipocytes are associated with intracellular membranes (Heller-Harrison, R., Morin, M. and Czech, M. (1995) J. Biol. Chem. 270, 24442-24450). We report here that in response to insulin, activated complexes of IRS-1.PI 3-kinase can be immunoprecipitated with anti-IRS-1 antibody from detergent extracts of immunoadsorbed GLUT4-containing vesicles prepared from 3T3-L1 adipocytes. The targeting of PI 3-kinase to rat adipocyte GLUT4-containing vesicles using vesicles prepared by sucrose velocity gradient ultracentrifugation was also demonstrated. Insulin treatment caused a 2.3-fold increase in immunoreactive p85 protein in these GLUT4-containing vesicles while anti-p85 immunoprecipitates of PI 3-kinase activity in GLUT4-containing vesicle extracts increased to a similar extent. HPLC analysis of the GLUT4 vesicle-associated PI 3-kinase activity showed insulin-mediated increases in PI 3-P, PI 3,4-P2, and PI 3,4,5-P3 when PI, PI 4-P, and PI 4,5-P2 were used as substrates. Our data demonstrate that insulin directs the association of PI 3-kinase with GLUT4-containing vesicles in 3T3-L1 and rat adipocytes, consistent with the hypothesis that PI 3-kinase is involved in the insulin-regulated movement of GLUT4 to the plasma membrane.
J Biol Chem. 1996 Apr 26;271(17):10200-4.
The Journal of biological chemistry
Heller-Harrison, Robin A.; Morin, Michelle; Guilherme, Adilson L.; and Czech, Michael P., "Insulin-mediated targeting of phosphatidylinositol 3-kinase to GLUT4-containing vesicles" (1996). Open Access Articles. 799.