Mechanism of activation of protein kinase B by insulin and IGF-1

Dario R. Alessi, Mirjana Andjelkovic, Barry Caudwell, Peter Cron, Nick Morrice, Philip Cohen, Brian A. Hemmings

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    Abstract

    Insulin activated endogenous protein kinase Ba (also known as RAC/Akt kinase) activity 12-fold in L6 myotubes, while after transfection into 293 cells PKBa was activated 20- and 50-fold in response to insulin and IGF-1 respectively. In both cells, the activation of PKBa was accompanied by its phosphorylation at Thr308 and Ser473 and, like activation, phosphorylation of both of these residues was prevented by the phosphatidylinositol 3-kinase inhibitor wortmannin. Thr308 and/or Ser473 were mutated to Ala or Asp and activities of mutant PKBa molecules were analysed after transfection into 293 cells. The activity of wild-type and mutant PKBa was also measured in vitro after stoichiometric phosphorylation of Ser473 by MAPKAP kinase-2. These experiments demonstrated that activation of PKBa by insulin or insulin-like growth factor-1 (IGF-1) results from phosphorylation of both Thr308 and Ser473, that phosphorylation of both residues is critical to generate a high level of PKBa activity and that the phosphorylation of Thr308 in vivo is not dependent on phosphorylation of Ser473 or vice versa. We propose a model whereby PKBa becomes phosphorylated and activated in insulin/IGF-1-stimulated cells by an upstream kinase(s).
    Original languageEnglish
    Pages (from-to)6541-6551
    Number of pages11
    JournalEMBO Journal
    Volume15
    Issue number23
    Publication statusPublished - 1996

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    Alessi, D. R., Andjelkovic, M., Caudwell, B., Cron, P., Morrice, N., Cohen, P., & Hemmings, B. A. (1996). Mechanism of activation of protein kinase B by insulin and IGF-1. EMBO Journal, 15(23), 6541-6551. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC452479/