title: Insulin stimulated phosphorylation of its own receptor. Activation of a tyrosine-specific protein kinase that is tightly associated with the receptor.
abstract: In solubilized, (wheat germ) lectin-purified preparations of rat liver membranes, insulin stimulated the incorporation of 32P from [gamma-32P]ATP into tyrosine residues of insulin receptor, casein, and histones. Despite the presence of both protein kinase and phosphatase activities in these preparations, no decrease in the 32P content of receptors (preincubated with or without insulin (0.5-100 nM)) was detected whether 32P incorporation was terminated by excess ATP, ATP + Mn2+, EDTA, or phosphatase inhibitors. Similarly, there was no decrease in the 32P content of phosphoreceptors incubated for up to 60 min with fresh receptor preparations in the presence or absence of insulin. Dephosphorylation of the insulin receptor to 20% of original 32P content only occurred when alkaline phosphatase was added to the preparations. It is concluded that endogenous receptor phosphatase(s) are either missing or inactive in these preparations, and consequently, insulin stimulates phosphorylation of its own receptor by activating a protein kinase. The kinase activity is tightly associated with the receptor itself; insulin also stimulated the phosphorylation of both receptor subunits in purified insulin-receptor complexes that had been immunoprecipitated by anti-insulin antibodies. However, the phosphorylating machinery is much more sensitive to heat inactivation than the binding function (90% less 32P incorporation versus 15% less binding during 60-min incubation at 37 degrees C), suggesting that the kinase is not associated exclusively with the insulin-binding domain.
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D004789:Enzyme Activation; D007328:Insulin / Q000494:pharmacology*; D007700:Kinetics; D008099:Liver / Q000378:metabolism; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism*; D011505:Protein-Tyrosine Kinases; D051381:Rats; D011972:Receptor, Insulin / Q000187:drug effects / Q000378:metabolism*; D013379:Substrate Specificity
chemical_list: D007328:Insulin; D011494:Protein Kinases; D011505:Protein-Tyrosine Kinases; D011972:Receptor, Insulin