Local frustration determines loop opening during the catalytic cycle of an oxidoreductase

  1. Lukas L Stelzl
  2. Despoina A I Mavridou
  3. Emmanuel Saridakis
  4. Diego Gonzalez
  5. Andrew J Baldwin
  6. Stuart J Ferguson
  7. Mark SP Sansom  Is a corresponding author
  8. Christina Redfield  Is a corresponding author
  1. University of Oxford, United Kingdom
  2. University of Texas at Austin, United States
  3. NCSR Demokritos, Greece
  4. University of Neuchatel, Switzerland

Abstract

Local structural frustration, the existence of mutually exclusive competing interactions, may explain why some proteins are dynamic while others are rigid. Frustration is thought to underpin biomolecular recognition and the flexibility of protein binding sites. Here we show how a small chemical modification, the oxidation of two cysteine thiols to a disulfide bond, during the catalytic cycle of the N-terminal domain of the key bacterial oxidoreductase DsbD (nDsbD), introduces frustration ultimately influencing protein function. In oxidized nDsbD, local frustration disrupts the packing of the protective cap-loop region against the active site allowing loop opening. By contrast, in reduced nDsbD the cap loop is rigid, always protecting the active-site thiols from the oxidizing environment of the periplasm. Our results point towards an intricate coupling between the dynamics of the active-site cysteines and of the cap loop which modulates the association reactions of nDsbD with its partners resulting in optimized protein function.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files.

The following data sets were generated
The following previously published data sets were used

Article and author information

Author details

  1. Lukas L Stelzl

    Department of Biochemistry, University of Oxford, Oxford, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  2. Despoina A I Mavridou

    Department of Molecular Biosciences, University of Texas at Austin, Austin, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Emmanuel Saridakis

    Institute of Nanoscience and Nanotechnology, NCSR Demokritos, Athens, Greece
    Competing interests
    The authors declare that no competing interests exist.
  4. Diego Gonzalez

    Department of Biology, University of Neuchatel, Neuchatel, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
  5. Andrew J Baldwin

    Physical and Theoretical Chemistry, University of Oxford, Oxford, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7579-8844
  6. Stuart J Ferguson

    Department of Biochemistry, University of Oxford, Oxford, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  7. Mark SP Sansom

    Department of Biochemistry, University of Oxford, Oxford, United Kingdom
    For correspondence
    mark.sansom@bioch.ox.ac.uk
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6360-7959
  8. Christina Redfield

    Department of Biochemistry, University of Oxford, Oxford, United Kingdom
    For correspondence
    christina.redfield@bioch.ox.ac.uk
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7297-7708

Funding

Biotechnology and Biological Sciences Research Council (BB/F01709X/1)

  • Lukas L Stelzl

Biotechnology and Biological Sciences Research Council (BB/R00126X/1)

  • Mark SP Sansom

Wellcome (208361/Z/17/Z)

  • Mark SP Sansom

Wellcome (079440)

  • Christina Redfield

Wellcome (092532/Z/10/Z)

  • Stuart J Ferguson
  • Christina Redfield

Swiss National Science Foundation (P300PA_167703)

  • Diego Gonzalez

Swiss National Science Foundation (PZ00P3_180142)

  • Diego Gonzalez

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

Reviewing Editor

  1. Lewis E Kay, University of Toronto, Canada

Publication history

  1. Received: December 21, 2019
  2. Accepted: June 21, 2020
  3. Accepted Manuscript published: June 22, 2020 (version 1)
  4. Version of Record published: July 9, 2020 (version 2)

Copyright

© 2020, Stelzl 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.

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  1. Lukas L Stelzl
  2. Despoina A I Mavridou
  3. Emmanuel Saridakis
  4. Diego Gonzalez
  5. Andrew J Baldwin
  6. Stuart J Ferguson
  7. Mark SP Sansom
  8. Christina Redfield
(2020)
Local frustration determines loop opening during the catalytic cycle of an oxidoreductase
eLife 9:e54661.
https://doi.org/10.7554/eLife.54661

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