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Identification of the Amino Acids 300-600 of IRS-2 as 14-3-3 Binding Region with the Importance of IGF-1/Insulin-Regulated Phosphorylation of Ser-573

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Identification of the Amino Acids 300-600 of IRS-2 as 14-3-3 Binding Region with the Importance of IGF-1/Insulin-Regulated Phosphorylation of Ser-573. / Neukamm, Sabine S; Toth, Rachel; Morrice, Nick; Campbell, David G; Mackintosh, Carol; Lehmann, Rainer; Haering, Hans-Ulrich; Schleicher, Erwin D; Weigert, Cora.

In: PLoS ONE, Vol. 7, No. 8, 2012, p. e43296.

Research output: Contribution to journalArticle

Harvard

Neukamm, SS, Toth, R, Morrice, N, Campbell, DG, Mackintosh, C, Lehmann, R, Haering, H-U, Schleicher, ED & Weigert, C 2012, 'Identification of the Amino Acids 300-600 of IRS-2 as 14-3-3 Binding Region with the Importance of IGF-1/Insulin-Regulated Phosphorylation of Ser-573' PLoS ONE, vol 7, no. 8, pp. e43296., 10.1371/journal.pone.0043296

APA

Neukamm, S. S., Toth, R., Morrice, N., Campbell, D. G., Mackintosh, C., Lehmann, R., ... Weigert, C. (2012). Identification of the Amino Acids 300-600 of IRS-2 as 14-3-3 Binding Region with the Importance of IGF-1/Insulin-Regulated Phosphorylation of Ser-573. PLoS ONE, 7(8), e43296. 10.1371/journal.pone.0043296

Vancouver

Neukamm SS, Toth R, Morrice N, Campbell DG, Mackintosh C, Lehmann R et al. Identification of the Amino Acids 300-600 of IRS-2 as 14-3-3 Binding Region with the Importance of IGF-1/Insulin-Regulated Phosphorylation of Ser-573. PLoS ONE. 2012;7(8):e43296. Available from: 10.1371/journal.pone.0043296

Author

Neukamm, Sabine S; Toth, Rachel; Morrice, Nick; Campbell, David G; Mackintosh, Carol; Lehmann, Rainer; Haering, Hans-Ulrich; Schleicher, Erwin D; Weigert, Cora / Identification of the Amino Acids 300-600 of IRS-2 as 14-3-3 Binding Region with the Importance of IGF-1/Insulin-Regulated Phosphorylation of Ser-573.

In: PLoS ONE, Vol. 7, No. 8, 2012, p. e43296.

Research output: Contribution to journalArticle

Bibtex - Download

@article{2daab50d0b6440249f080bcbc9d6c8e4,
title = "Identification of the Amino Acids 300-600 of IRS-2 as 14-3-3 Binding Region with the Importance of IGF-1/Insulin-Regulated Phosphorylation of Ser-573",
author = "Neukamm, {Sabine S} and Rachel Toth and Nick Morrice and Campbell, {David G} and Carol Mackintosh and Rainer Lehmann and Hans-Ulrich Haering and Schleicher, {Erwin D} and Cora Weigert",
year = "2012",
doi = "10.1371/journal.pone.0043296",
volume = "7",
number = "8",
pages = "e43296",
journal = "PLoS ONE",

}

RIS (suitable for import to EndNote) - Download

TY - JOUR

T1 - Identification of the Amino Acids 300-600 of IRS-2 as 14-3-3 Binding Region with the Importance of IGF-1/Insulin-Regulated Phosphorylation of Ser-573

A1 - Neukamm,Sabine S

A1 - Toth,Rachel

A1 - Morrice,Nick

A1 - Campbell,David G

A1 - Mackintosh,Carol

A1 - Lehmann,Rainer

A1 - Haering,Hans-Ulrich

A1 - Schleicher,Erwin D

A1 - Weigert,Cora

AU - Neukamm,Sabine S

AU - Toth,Rachel

AU - Morrice,Nick

AU - Campbell,David G

AU - Mackintosh,Carol

AU - Lehmann,Rainer

AU - Haering,Hans-Ulrich

AU - Schleicher,Erwin D

AU - Weigert,Cora

PY - 2012

Y1 - 2012

N2 - Phosphorylation of insulin receptor substrate (IRS)-2 on tyrosine residues is a key event in IGF-1/insulin signaling and leads to activation of the PI 3-kinase and the Ras/MAPK pathway. Furthermore, phosphorylated serine/threonine residues on IRS-2 can induce 14-3-3 binding. In this study we searched IRS-2 for novel phosphorylation sites and investigated the interaction between IRS-2 and 14-3-3. Mass spectrometry identified a total of 24 serine/threonine residues on IRS-2 with 12 sites unique for IRS-2 while the other residues are conserved in IRS-1 and IRS-2. IGF-1 stimulation led to increased binding of 14-3-3 to IRS-2 in transfected HEK293 cells and this binding was prevented by inhibition of the PI 3-kinase pathway and an Akt/PKB inhibitor. Insulin-stimulated interaction between endogenous IRS-2 and 14-3-3 was observed in rat hepatoma cells and in mice liver after an acute insulin stimulus and refeeding. Using different IRS-2 fragments enabled localization of the IGF-1-dependent 14-3-3 binding region spanning amino acids 300-600. The 24 identified residues on IRS-2 included several 14-3-3 binding candidates in the region 300-600. Single alanine mutants of these candidates led to the identification of serine 573 as 14-3-3 binding site. A phospho-site specific antibody was generated to further characterize serine 573. IGF-1-dependent phosphorylation of serine 573 was reduced by inhibition of PI 3-kinase and Akt/PKB. A negative role of this phosphorylation site was implicated by the alanine mutant of serine 573 which led to enhanced phosphorylation of Akt/PKB in an IGF-1 time course experiment. To conclude, our data suggest a physiologically relevant role for IGF-1/insulin-dependent 14-3-3 binding to IRS-2 involving serine 573.

AB - Phosphorylation of insulin receptor substrate (IRS)-2 on tyrosine residues is a key event in IGF-1/insulin signaling and leads to activation of the PI 3-kinase and the Ras/MAPK pathway. Furthermore, phosphorylated serine/threonine residues on IRS-2 can induce 14-3-3 binding. In this study we searched IRS-2 for novel phosphorylation sites and investigated the interaction between IRS-2 and 14-3-3. Mass spectrometry identified a total of 24 serine/threonine residues on IRS-2 with 12 sites unique for IRS-2 while the other residues are conserved in IRS-1 and IRS-2. IGF-1 stimulation led to increased binding of 14-3-3 to IRS-2 in transfected HEK293 cells and this binding was prevented by inhibition of the PI 3-kinase pathway and an Akt/PKB inhibitor. Insulin-stimulated interaction between endogenous IRS-2 and 14-3-3 was observed in rat hepatoma cells and in mice liver after an acute insulin stimulus and refeeding. Using different IRS-2 fragments enabled localization of the IGF-1-dependent 14-3-3 binding region spanning amino acids 300-600. The 24 identified residues on IRS-2 included several 14-3-3 binding candidates in the region 300-600. Single alanine mutants of these candidates led to the identification of serine 573 as 14-3-3 binding site. A phospho-site specific antibody was generated to further characterize serine 573. IGF-1-dependent phosphorylation of serine 573 was reduced by inhibition of PI 3-kinase and Akt/PKB. A negative role of this phosphorylation site was implicated by the alanine mutant of serine 573 which led to enhanced phosphorylation of Akt/PKB in an IGF-1 time course experiment. To conclude, our data suggest a physiologically relevant role for IGF-1/insulin-dependent 14-3-3 binding to IRS-2 involving serine 573.

U2 - 10.1371/journal.pone.0043296

DO - 10.1371/journal.pone.0043296

M1 - Article

JO - PLoS ONE

JF - PLoS ONE

IS - 8

VL - 7

SP - e43296

ER -

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