Activation of phosphatidylinositol 3-kinase by insulin.
Insulin action appears to require the protein-tyrosine kinase domain of the beta subunit of the insulin receptor. Despite this, the identities and biochemical functions of the cellular targets of this tyrosine kinase are unknown. A phosphatidylinositol 3-kinase (PI 3-kinase) that phosphorylates the D-3 position of the inositol ring associates with several protein-tyrosine kinases. Here we report that PI 3-kinase activity is immunoprecipitated from insulin-stimulated CHO cells by antiphosphotyrosine and anti-insulin receptor antibodies. Insulin as low as 0.3 nM increased immunoprecipitable PI 3-kinase activity within 1 min. Increases in activity were much greater in CHO cells expressing the human insulin receptor (100,000 receptors per cell) than in control CHO cells (2000 receptors per cell). During insulin stimulation, various lipid products of the PI 3-kinase either appeared or increased in quantity in intact cells, suggesting that the appearance of immunoprecipitable PI 3-kinase reflects an increase in its activity in vivo. These results indicate that insulin at physiological concentrations regulates the PI 3-kinase and suggest that this regulation involves a physical association between the insulin receptor and the PI 3-kinase and tyrosyl phosphorylation.
Ruderman
N B
NB
Department of Physiology, Tufts University School of Medicine, Boston, MA 02111.
Kapeller
R
R
White
M F
MF
Cantley
L C
LC
D000818:Animals; D000906:Antibodies; D002460:Cell Line; D004789:Enzyme Activation; D007328:Insulin / Q000494:pharmacology*; D007700:Kinetics; D019869:Phosphatidylinositol 3-Kinases; D010716:Phosphatidylinositols / Q000096:biosynthesis / Q000302:isolation & purification; D010770:Phosphotransferases / Q000378:metabolism*; D011972:Receptor, Insulin / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D014162:Transfection
D000906:Antibodies; D007328:Insulin; D010716:Phosphatidylinositols; D010770:Phosphotransferases; D019869:Phosphatidylinositol 3-Kinases; D011972:Receptor, Insulin