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Heart 6-phosphofructo-2-kinase activation by insulin requires PKB (protein kinase B), but not SGK3 (serum- and glucocorticoid-induced protein kinase 3).

Mouton, Véronique and Toussaint, Louise and Vertommen, Didier and Gueuning, Marie-Agnès and Maisin, Liliane and Havaux, Xavier and Sanchez-Canedo, Cossette and Bertrand, Luc and Dequiedt, Franck and Hemmings, Brian Arthur and Hue, Louis and Rider, Mark H (2010) Heart 6-phosphofructo-2-kinase activation by insulin requires PKB (protein kinase B), but not SGK3 (serum- and glucocorticoid-induced protein kinase 3). The Biochemical Journal, 431 (2). pp. 267-275. ISSN 1470-8728

Abstract

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.

Item Type: Article
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Additional Information: unclear
Keywords: glycolysis; protein kinase B (PKB); 6-phosphofructo-2-kinase (PFK-2); serum- and glucocorticoid-induced protein kinase 3 (SGK3); small interfering RNA (siRNA)
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Date Deposited: 04 Dec 2010 00:45
Last Modified: 01 Feb 2013 00:48
URI: https://oak.novartis.com/id/eprint/3907

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