Analysis of insulin-receptor phosphorylation sites in intact cells by two-dimensional phosphopeptide mapping.

作者: K Siddle , R M Denton , J M Tavaré , R M O'Brien

DOI: 10.1042/BJ2530783

关键词: AutophosphorylationProtein tyrosine phosphataseBiochemistryTyrosineInsulin receptorIRS2Receptor tyrosine kinaseSH2 domainMolecular biologyBiologyPhosphorylation

摘要: Insulin stimulates the autophosphorylation of partially purified insulin receptor initially on tyrosine residues 1146, 1150 and 1151. This is followed by increased 1316, 1322 two further residues, possibly 953 960 or 972 [Tavare & Denton (1988) Biochem. J. 252, 607-615]. In present paper we have used cell lines transfected with insulin-receptor cDNA (CHO.T NIH 3T3 HIR3.5 cells) to assess which are phosphorylated within intact cells. We show that: (1) causes a rapid increase in phosphorylation 1151 both types; 1316 also phosphorylated, but apparently lesser extent cells; (2) sites that may correspond appear be very poorly (3) promotes substantial serine threonine receptors CHO.T this results appearance phosphopeptides not evident maps solubilized preparations autophosphorylated vitro.

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