Grb10 inhibits insulin-stimulated insulin receptor substrate (IRS)-phosphatidylinositol 3-kinase/Akt signaling pathway by disrupting the association of IRS-1/IRS-2 with the insulin receptor

KeriLyn R. Wick, Eric D. Werner, Paul Langlais, Fresnida J. Ramos, Lily Q Dong, Steven E. Shoelson, Feng Liu

Research output: Contribution to journalArticle

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Abstract

Grb10 has been proposed to inhibit or activate insulin signaling, depending on cellular context. We have investigated the mechanism by which full-length hGrb10γ inhibits signaling through the insulin receptor substrate (IRS) proteins. Overexpression of hGrb10γ in CHO/IR cells and in differentiated adipocytes significantly reduced insulin-stimulated tyrosine phosphorylation of IRS-1 and IRS-2. Inhibition occurred rapidly and was sustained for 60 min during insulin stimulation. In agreement with inhibited signaling through the IRS/PI 3-kinase pathway, we found hGrb10γ to both delay and reduce phosphorylation of Akt at Thr308 and Ser473 in response to insulin stimulation. Decreased phosphorylation of IRS-1/2 may arise from impaired catalytic activity of the receptor, since hGrb10γ directly associates with the IR kinase regulatory loop. However, yeast tri-hybrid studies indicated that full-length Grb10 blocks association between IRS proteins and IR, and that this requires the SH2 domain of Grb10. In cells, hGrb10γ inhibited insulin-stimulated IRS-1 tyrosine phosphorylation in a dose-dependent manner, but did not affect IR catalytic activity toward Tyr972 in the juxtamembrane region and Tyr1158/1162/1163 in the regulatory domain. We conclude that binding of hGrb10γ to IR decreases signaling through the IRS/PI 3-kinase/AKT pathway by physically blocking IRS access to IR.

Original languageEnglish (US)
Pages (from-to)8460-8467
Number of pages8
JournalJournal of Biological Chemistry
Volume278
Issue number10
DOIs
StatePublished - Mar 7 2003

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Phosphatidylinositol 3-Kinase
Insulin Receptor Substrate Proteins
Insulin Receptor
Phosphorylation
Association reactions
Insulin
Substrates
Phosphatidylinositol 3-Kinases
Tyrosine
Catalyst activity
Yeast
Phosphotransferases
src Homology Domains
CHO Cells
Adipocytes
Yeasts

ASJC Scopus subject areas

  • Biochemistry

Cite this

Grb10 inhibits insulin-stimulated insulin receptor substrate (IRS)-phosphatidylinositol 3-kinase/Akt signaling pathway by disrupting the association of IRS-1/IRS-2 with the insulin receptor. / Wick, KeriLyn R.; Werner, Eric D.; Langlais, Paul; Ramos, Fresnida J.; Dong, Lily Q; Shoelson, Steven E.; Liu, Feng.

In: Journal of Biological Chemistry, Vol. 278, No. 10, 07.03.2003, p. 8460-8467.

Research output: Contribution to journalArticle

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AU - Werner, Eric D.

AU - Langlais, Paul

AU - Ramos, Fresnida J.

AU - Dong, Lily Q

AU - Shoelson, Steven E.

AU - Liu, Feng

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