1. Biochemistry and Chemical Biology
  2. Structural Biology and Molecular Biophysics

PH domain-mediated autoinhibition and oncogenic activation of Akt

  1. Hwan Bae
  2. Thibault Viennet
  3. Eunyoung Park
  4. Nam Chu
  5. Antonieta Salguero
  6. Michael J Eck  Is a corresponding author
  7. Haribabu Arthanari  Is a corresponding author
  8. Philip A Cole  Is a corresponding author
  1. Harvard Medical School, United States
  2. Dana-Farber Cancer Institute, United States
  3. The Ohio State University, United States
  4. Brigham and Women's Hospital, United States
https://doi.org/10.7554/eLife.80148
Share this article
Cite this article
  1. Hwan Bae
  2. Thibault Viennet
  3. Eunyoung Park
  4. Nam Chu
  5. Antonieta Salguero
  6. Michael J Eck
  7. Haribabu Arthanari
  8. Philip A Cole
(2022)
PH domain-mediated autoinhibition and oncogenic activation of Akt
eLife 11:e80148.
https://doi.org/10.7554/eLife.80148
  2. Download BibTeX
  3. Download .RIS

Abstract

Akt is a Ser/Thr protein kinase that plays a central role in metabolism and cancer. Regulation of Akt's activity involves an autoinhibitory intramolecular interaction between its pleckstrin homology (PH) domain and its kinase domain that can be relieved by C-tail phosphorylation. PH domain mutant E17K Akt is a well-established oncogene. Previously, we reported that the conformation of autoinhibited Akt may be shifted by small molecule allosteric inhibitors limiting the mechanistic insights from existing X-ray structures that have relied on such compounds (Chu, Viennet, et al, 2020). Here we discover unexpectedly that a single mutation R86A Akt exhibits intensified autoinhibitory features with enhanced PH domain-kinase domain affinity. Structural and biochemical analysis uncovers the importance of a key interaction network involving Arg86, Glu17, and Tyr18 that controls Akt conformation and activity. Our studies also shed light on the molecular basis for E17K Akt activation as an oncogenic driver.

Data availability

Diffraction data have been deposited in PDB under the accession code: 7MYXSource data are uploaded as Zip files on the eLife website

Article and author information

Author details

  1. Hwan Bae

    Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5252-252X

  2. Thibault Viennet

    Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5349-0179

  3. Eunyoung Park

    Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5618-1267

  4. Nam Chu

    Department of Cancer Biology and Genetics, The Ohio State University, Columbus, United States
    Competing interests
    No competing interests declared.

  5. Antonieta Salguero

    Department of Medicine, Brigham and Women's Hospital, Boston, United States
    Competing interests
    No competing interests declared.

  6. Michael J Eck

    Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, United States
    For correspondence
    eck@crystal.harvard.edu
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-4247-9403

  7. Haribabu Arthanari

    Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, United States
    For correspondence
    hari_arthanari@hms.harvard.edu
    Competing interests
    No competing interests declared.

  8. Philip A Cole

    Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, United States
    For correspondence
    pacole@bwh.harvard.edu
    Competing interests
    Philip A Cole, Senior editor, eLifePhilip Cole has been a paid consultant for Scorpion Therapeutics..
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6873-7824

Funding

National Cancer Institute (R01CA74305)

  • Philip A Cole

National Cancer Institute (R35CA242461)

  • Michael J Eck

National Cancer Institute (K22CA241105)

  • Nam Chu

Claudia Adams Barr Program (Grant)

  • Haribabu Arthanari

National Institutes of Health (EB002026)

  • Haribabu Arthanari

Kwanjeong Education Foundation (Fellowship)

  • Hwan Bae

National Institute of General Medical Sciences (GM124165)

  • Michael J Eck

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Copyright

© 2022, Bae et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 2,987
    views
  • 721
    downloads
  • 18
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Hwan Bae
  2. Thibault Viennet
  3. Eunyoung Park
  4. Nam Chu
  5. Antonieta Salguero
  6. Michael J Eck
  7. Haribabu Arthanari
  8. Philip A Cole
(2022)
PH domain-mediated autoinhibition and oncogenic activation of Akt
eLife 11:e80148.
https://doi.org/10.7554/eLife.80148

Share this article

https://doi.org/10.7554/eLife.80148

Categories and tags

Research organism

Further reading

    1. Biochemistry and Chemical Biology

    The structural determinants of PH domain-mediated regulation of Akt revealed by segmental labeling

    Nam Chu, Thibault Viennet ... Philip A Cole
    Research Article Updated

    Akt is a critical protein kinase that governs cancer cell growth and metabolism. Akt appears to be autoinhibited by an intramolecular interaction between its N-terminal pleckstrin homology (PH) domain and kinase domain, which is relieved by C-tail phosphorylation, but the precise molecular mechanisms remain elusive. Here, we use a combination of protein semisynthesis, NMR, and enzymological analysis to characterize structural features of the PH domain in its autoinhibited and activated states. We find that Akt autoinhibition depends on the length/flexibility of the PH-kinase linker. We identify a role for a dynamic short segment in the PH domain that appears to regulate autoinhibition and PDK1-catalyzed phosphorylation of Thr308 in the activation loop. We determine that Akt allosteric inhibitor MK2206 drives distinct PH domain structural changes compared to baseline autoinhibited Akt. These results highlight how the conformational plasticity of Akt governs the delicate control of its catalytic properties.

    1. Biochemistry and Chemical Biology
    2. Structural Biology and Molecular Biophysics
    3. Cell Biology

    Special Issue: Allosteric Regulation of Kinase Activity

    Edited by Volker Dötsch et al.
    Collection

    Our latest Special Issue brings together research on the allosteric regulation of kinase activity.

    1. Biochemistry and Chemical Biology

    Enzymatic protein fusions with 100% product yield

    Adrian CD Fuchs
    Research Article

    The protein ligase Connectase can be used to fuse proteins to small molecules, solid carriers, or other proteins. Compared to other protein ligases, it offers greater substrate specificity, higher catalytic efficiency, and catalyzes no side reactions. However, its reaction is reversible, resulting in only 50% fusion product from two equally abundant educts. Here, we present a simple method to reliably obtain 100% fusion product in 1:1 conjugation reactions. This method is efficient for protein-protein or protein-peptide fusions at the N- or C-termini. It enables the generation of defined and completely labeled antibody conjugates with one fusion partner on each chain. The reaction requires short incubation times with small amounts of enzyme and is effective even at low substrate concentrations and at low temperatures. With these characteristics, it presents a valuable new tool for bioengineering.