title: The beta subunit of the insulin receptor is an insulin-activated protein kinase.
abstract: In the presence of adenosine 5'-[gamma-32P]triphosphate ([gamma-32P]ATP) and a partially purified human placental insulin receptor preparation, insulin stimulates the phosphorylation of an Mr 94000 protein in a time- and dose-dependent manner. Half-maximal stimulation of 32P incorporation occurs at (2-3) X 10(-9) M insulin, a concentration identical with the Kd for insulin binding in this preparation. Immunoprecipitations with monoclonal anti-insulin receptor antibody demonstrate that the Mr 94000 protein kinase substrate is a component of the insulin receptor, the beta subunit. If the partially purified, soluble placental receptor preparation is immunoprecipitated and then exposed to [gamma-32P]ATP and insulin, phosphorylation of the Mr 94000 protein is maintained. The photoincorporation of 8-azido[alpha-32P]ATP into placental insulin receptor preparations was carried out to identify the ATP binding site responsible for the protein kinase activity. Photoincorporation into numerous proteins was observed, including both subunits of the insulin receptor. However, when photolabeling was performed in the presence of excess adenosine 5'-(beta, gamma-imidotriphosphate), a nonhydrolyzable ATP derivative, the beta subunit of the insulin receptor was the only species protected from label incorporation. These data indicate that the beta subunit of the insulin receptor has insulin-dependent protein kinase activity. Phosphotyrosine formation is the primary result of this activity in placental insulin receptor preparations.
mesh_terms: D005260:Female; D006801:Humans; D007328:Insulin / Q000494:pharmacology*; D007700:Kinetics; D046911:Macromolecular Substances; D010920:Placenta / Q000378:metabolism*; D011247:Pregnancy; D011494:Protein Kinases / Q000302:isolation & purification* / Q000378:metabolism; D011972:Receptor, Insulin / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D007328:Insulin; D046911:Macromolecular Substances; D011494:Protein Kinases; D011972:Receptor, Insulin