Insulin-mediated targeting of phosphatidylinositol 3-kinase to GLUT4-containing vesicles
UMass Chan Affiliations
Department of Biochemistry and Molecular BiologyProgram in Molecular Medicine
Document Type
Journal ArticlePublication Date
1996-04-26Keywords
1-Phosphatidylinositol 3-Kinase3T3 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
Metadata
Show full item recordAbstract
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.Source
J Biol Chem. 1996 Apr 26;271(17):10200-4.
DOI
10.1074/jbc.271.17.10200Permanent Link to this Item
http://hdl.handle.net/20.500.14038/42450PubMed ID
8626583Related Resources
ae974a485f413a2113503eed53cd6c53
10.1074/jbc.271.17.10200