On the basis of transfection experiments using a dominant-negative approach, our previous studies suggested that PKB (protein kinase B) was not involved in heart PFK-2 (6-phosphofructo2-kinase) activation by insulin. Therefore we first tested whether SGK3 (serum- and glucocorticoid-induced protein kinase 3) might be involved in this effect. Treatment of recombinant heart PFK-2 with [γ-32P]ATP and SGK3 in vitro led to PFK-2 activation and phosphorylation at Ser466 and Ser483. However, in HEK-293T cells [HEK (human embryonic kidney)-293 cells expressing the large T-antigen of SV40 (simian virus 40)] co-transfected with SGK3 siRNA (small interfering RNA) and heart PFK-2, insulin-induced heart PFK-2 activation was unaffected. The involvement of PKB in heart PFK-2 activation by insulin was re-evaluated using different models: (i) hearts from transgenic mice with a muscle/heart-specific mutation in the PDK1 (phosphoinositide-dependent protein kinase 1)-substrate-docking site injected with insulin; (ii) hearts from PKBβ-deficient mice injected with insulin; (iii) freshly isolated rat cardiomyocytes and perfused hearts treated with the selective Akti-1/2 PKB inhibitor prior to insulin treatment; and (iv) HEK-293T cells co-transfected with heart PFK-2, and PKBα/β siRNA or PKBα siRNA, incubated with insulin. Together, the results indicated that SGK3 is not required for insulin-induced PFK-2 activation and that this effect is likely mediated by PKBα.
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Research Article|
September 28 2010
Heart 6-phosphofructo-2-kinase activation by insulin requires PKB (protein kinase B), but not SGK3 (serum- and glucocorticoid-induced protein kinase 3)
Véronique Mouton;
Véronique Mouton
1
*Université catholique de Louvain and de Duve Institute, 75 Avenue Hippocrate, B-1200 Brussels, Belgium
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Louise Toussaint;
Louise Toussaint
1
*Université catholique de Louvain and de Duve Institute, 75 Avenue Hippocrate, B-1200 Brussels, Belgium
†Cellular and Molecular Biology Unit, FUSAGx, 13 Avenue Marechal Juin, 5030 Gembloux, Belgium
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Didier Vertommen;
Didier Vertommen
*Université catholique de Louvain and de Duve Institute, 75 Avenue Hippocrate, B-1200 Brussels, Belgium
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Marie-Agnès Gueuning;
Marie-Agnès Gueuning
*Université catholique de Louvain and de Duve Institute, 75 Avenue Hippocrate, B-1200 Brussels, Belgium
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Liliane Maisin;
Liliane Maisin
*Université catholique de Louvain and de Duve Institute, 75 Avenue Hippocrate, B-1200 Brussels, Belgium
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Xavier Havaux;
Xavier Havaux
‡Université catholique de Louvain Division of Cardiology, 55 Avenue Hippocrate, CARD5550, B-1200 Brussels, Belgium
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Cossette Sanchez-Canedo;
Cossette Sanchez-Canedo
‡Université catholique de Louvain Division of Cardiology, 55 Avenue Hippocrate, CARD5550, B-1200 Brussels, Belgium
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Luc Bertrand;
Luc Bertrand
‡Université catholique de Louvain Division of Cardiology, 55 Avenue Hippocrate, CARD5550, B-1200 Brussels, Belgium
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Franck Dequiedt;
Franck Dequiedt
†Cellular and Molecular Biology Unit, FUSAGx, 13 Avenue Marechal Juin, 5030 Gembloux, Belgium
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Brian A. Hemmings;
Brian A. Hemmings
§Friedrich Miescher Institute for Biomedical Research, Maulbeerstrasse 66, 4058 Basel, Switzerland
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Louis Hue;
Louis Hue
*Université catholique de Louvain and de Duve Institute, 75 Avenue Hippocrate, B-1200 Brussels, Belgium
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Mark H. Rider
Mark H. Rider
2
*Université catholique de Louvain and de Duve Institute, 75 Avenue Hippocrate, B-1200 Brussels, Belgium
2To whom correspondence should be addressed (email mark.rider@uclouvain.be).
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Publisher: Portland Press Ltd
Received:
July 19 2010
Accepted:
August 05 2010
Accepted Manuscript online:
August 05 2010
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© The Authors Journal compilation © 2010 Biochemical Society
2010
Biochem J (2010) 431 (2): 267–275.
Article history
Received:
July 19 2010
Accepted:
August 05 2010
Accepted Manuscript online:
August 05 2010
Citation
Véronique Mouton, Louise Toussaint, Didier Vertommen, Marie-Agnès Gueuning, Liliane Maisin, Xavier Havaux, Cossette Sanchez-Canedo, Luc Bertrand, Franck Dequiedt, Brian A. Hemmings, Louis Hue, Mark H. Rider; Heart 6-phosphofructo-2-kinase activation by insulin requires PKB (protein kinase B), but not SGK3 (serum- and glucocorticoid-induced protein kinase 3). Biochem J 15 October 2010; 431 (2): 267–275. doi: https://doi.org/10.1042/BJ20101089
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