Insulin stimulation of phosphatidylinositol 3-kinase activity and association with insulin receptor substrate 1 in liver and muscle of the intact rat

Franco Folli, Mario J A Saad, Jonathan M. Backer, C. Ronald Kahn

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Abstract

Growth factors stimulate the enzyme phosphatidylinositol (PI) 3-kinase in cells in culture. Insulin rapidly stimulates tyrosine phosphorylation of its endogenous substrate, insulin receptor substrate 1 (IRS-1), and in vitro IRS-1 associates with PI 3-kinase, thus activating the enzyme. We have examined whether insulin is capable of stimulating the PI 3-kinase pathway in two physiological target tissues for the actions of insulin in vivo, liver and skeletal muscle. After intraportal injection of insulin into anesthetized rats, there was a 2-fold stimulation of total hepatic PI 3-kinase activity in liver and muscle extracts and a 10- to 20-fold increase in PI 3-kinase activity immunoprecipitated with anti-IRS-1 antibodies. Stimulation of PI 3-kinase was accompanied by an association between this enzyme and IRS-I as detected by immunoprecipitation of liver and muscle extracts with anti-IRS-1 antibodies and Western blotting with antibodies to the 85-kDa subunit of PI 3-kinase. Immunoprecipitation with anti-p85 antibodies and phosphotyrosine immunoblotting revealed no tyrosine phosphorylation of PI 3-kinase, but demonstrated co-precipitation of tyrosine-phosphorylated IRS-1, as well as another phosphotyrosine protein of ∼135-140 kDa. Thus, IRS-1 phosphorylation plays a significant role in the activation of PI 3-kinase in vivo by insulin.

Original languageEnglish (US)
Pages (from-to)22171-22177
Number of pages7
JournalJournal of Biological Chemistry
Volume267
Issue number31
StatePublished - Nov 5 1992
Externally publishedYes

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Phosphatidylinositol 3-Kinase
Insulin Receptor Substrate Proteins
Liver
Muscle
Rats
Association reactions
Insulin
Muscles
Phosphorylation
Tyrosine
Liver Extracts
Phosphotyrosine
Antibodies
Immunoprecipitation
Enzymes
Coprecipitation
Immunoblotting
Anti-Idiotypic Antibodies
Intercellular Signaling Peptides and Proteins
Skeletal Muscle

ASJC Scopus subject areas

  • Biochemistry

Cite this

Insulin stimulation of phosphatidylinositol 3-kinase activity and association with insulin receptor substrate 1 in liver and muscle of the intact rat. / Folli, Franco; Saad, Mario J A; Backer, Jonathan M.; Kahn, C. Ronald.

In: Journal of Biological Chemistry, Vol. 267, No. 31, 05.11.1992, p. 22171-22177.

Research output: Contribution to journalArticle

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