Direct activation of the phosphatidylinositol 3′-kinase by the insulin receptor

Debra J. Van Horn, Martin G. Myers, Jonathan M. Backer

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Abstract

We have previously shown that phosphatidylinositol (PtdIns) 3′-kinase is activated by the binding of proteins or peptides containing the phosphorylated motif Y(P)XXM. In the present study, we examine interactions between Ptdlns 3′-kinase and the human insulin receptor, which contains a C-terminal phosphorylation site in the sequence Y1322THM. Partially purified insulin receptors bound tightly to bacterial fusion proteins containing the N- or C-terminal SH2 domains from Ptdlns 3′-kinase regulatory subunit (p85). In contrast, a mutant insulin receptor, truncated by 43 amino acids at the C terminus (IRΔCT), bound poorly to the SH2 domains; these mutant receptors have normal kinase activity but lack the Y1322THM motif. Similarly, incubation with wild-type receptors increased the activity of immunopurified Ptdlns 3′-kinase, whereas incubation with IRΔCT receptors did not affect Ptdlns 3′-kinase activity. Activation of Ptdlns 3′-kinase by the wild-type receptor was mimicked by a tyrosyl phosphopeptide derived from the insulin receptor C terminus and containing the Y1322THM motif; non-phosphorylated peptide did not affect activity. Thus, the insulin receptor C terminus activates Ptdlns 3′-kinase in vitro by binding to the SH2 domains of the 85-kDa regulatory subunit. These data support the hypothesis that binding of tyrosyl-phosphorylated receptors to p85 SH2 domains is a general mechanism for Ptdlns 3′-kinase activation, and they suggest that direct interactions between the insulin receptor and Ptdlns 3′-kinase may provide an alternative pathway for the activation of this enzyme by insulin.

Original languageEnglish (US)
Pages (from-to)29-32
Number of pages4
JournalJournal of Biological Chemistry
Volume269
Issue number1
StatePublished - Jan 7 1994

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Phosphatidylinositol 3-Kinase
Insulin Receptor
Phosphotransferases
Chemical activation
src Homology Domains
Phosphopeptides
Peptides
Phosphorylation
Enzyme Activation
Bacterial Proteins
Carrier Proteins
Fusion reactions
Insulin
Amino Acids

ASJC Scopus subject areas

  • Biochemistry

Cite this

Direct activation of the phosphatidylinositol 3′-kinase by the insulin receptor. / Van Horn, Debra J.; Myers, Martin G.; Backer, Jonathan M.

In: Journal of Biological Chemistry, Vol. 269, No. 1, 07.01.1994, p. 29-32.

Research output: Contribution to journalArticle

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