Crystal structure of an activated Akt/Protein Kinase B ternary complex with GSK3-peptide and AMP-PNP (original) (raw)

Nature Structural Biology volume 9, pages 940–944 (2002)Cite this article

Abstract

The protein kinase Akt/PKB is stimulated by the phosphorylation of two regulatory residues, Thr 309 of the activation segment and Ser 474 of the hydrophobic motif (HM), that are structurally and functionally conserved within the AGC kinase family. To understand the mechanism of PKB regulation, we determined the crystal structures of activated kinase domains of PKB in complex with a GSK3β-peptide substrate and an ATP analog. The activated state of the kinase was generated by phosphorylating Thr 309 using PDK1 and mimicking Ser 474 phosphorylation either with the S474D substitution or by replacing the HM of PKB with that of PIFtide, a potent mimic of a phosphorylated HM. Comparison with the inactive PKB structure indicates that the role of Ser 474 phosphorylation is to promote the engagement of the HM with the N-lobe of the kinase domain, promoting a disorder-to-order transition of the αC helix. The αC helix, by interacting with pThr 309, restructures and orders the activation segment, generating an active kinase conformation. Analysis of the interactions between PKB and the GSK3β-peptide explains how PKB selects for protein substrates distinct from those of PKA.

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Acknowledgements

We thank the staff at the ESRF, Grenoble, France, for access to synchrotron radiation facilities, and T. Dale (ICR) for the GSK3β peptide. The work was supported by grants from Cancer Research UK and from the ICR to D.B. Friedrich Miescher-Institut is part of the Novartis Research Foundation.

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Authors and Affiliations

  1. Section of Structural Biology, Institute of Cancer Research, Chester Beatty Laboratories, 237 Fulham Road, London, SW3 6JB, UK
    Jing Yang, Valerie M. Good, Vivienne Thompson & David Barford
  2. Friedrich Miescher-Institut, Maulbeerstrasse 66, Basel, CH-4048, Switzerland
    Peter Cron & Brian A. Hemmings

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  1. Jing Yang
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  2. Peter Cron
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  3. Valerie M. Good
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  4. Vivienne Thompson
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  5. Brian A. Hemmings
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  6. David Barford
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Correspondence toDavid Barford.

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Yang, J., Cron, P., Good, V. et al. Crystal structure of an activated Akt/Protein Kinase B ternary complex with GSK3-peptide and AMP-PNP.Nat Struct Mol Biol 9, 940–944 (2002). https://doi.org/10.1038/nsb870

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