1. Structural Biology and Molecular Biophysics
  2. Neuroscience
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Identification of a pre-active conformation of a pentameric channel receptor

  1. Anaïs Menny
  2. Solène N Lefebvre
  3. Philipp AM Schmidpeter
  4. Emmanuelle Drège
  5. Zaineb Fourati
  6. Marc Delarue
  7. Stuart J Edelstein
  8. Crina M Nimigean
  9. Delphine Joseph
  10. Pierre-Jean Corringer  Is a corresponding author
  1. Institut Pasteur, France
  2. Weill Cornell Medicine, United States
  3. Université Paris-Sud, France
  4. Inserm U1024, CNRS 8197, Institute of Biology, Ecole Normale Supérieure, France
Research Article
  • Cited 23
  • Views 1,775
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Cite this article as: eLife 2017;6:e23955 doi: 10.7554/eLife.23955

Abstract

Pentameric ligand-gated ion channels (pLGICs) mediate fast chemical signalling through global allosteric transitions. Despite the existence of several high-resolution structures of pLGICs, their dynamical properties remain elusive. Using the proton-gated channel GLIC, we engineered multiple fluorescent reporters, each incorporating a bimane and a tryptophan/tyrosine, whose close distance causes fluorescence quenching. We show that proton application causes a global compaction of the extracellular subunit interface, coupled to an outward motion of the M2-M3 loop near the channel gate. These movements are highly similar in lipid vesicles and detergent micelles. These reorganizations are essentially completed within 2 ms and occur without channel opening at low proton concentration, indicating that they report a pre-active intermediate state in the transition pathway towards activation. This provides a template to investigate the gating of eukaryotic neurotransmitter receptors, for which intermediate states also participate in activation.

Article and author information

Author details

  1. Anaïs Menny

    Channel Receptors Unit, Institut Pasteur, Paris, France
    Competing interests
    The authors declare that no competing interests exist.
  2. Solène N Lefebvre

    Channel Receptors Unit, Institut Pasteur, Paris, France
    Competing interests
    The authors declare that no competing interests exist.
  3. Philipp AM Schmidpeter

    Department of Anesthesiology, Weill Cornell Medicine, New York, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Emmanuelle Drège

    BioCIS, Université Paris-Sud, Châtenay-Malabry, France
    Competing interests
    The authors declare that no competing interests exist.
  5. Zaineb Fourati

    Unité de Dynamique Structurale des Macromolécules, Institut Pasteur, Paris, France
    Competing interests
    The authors declare that no competing interests exist.
  6. Marc Delarue

    Unité de Dynamique Structurale des Macromolécules, Institut Pasteur, Paris, France
    Competing interests
    The authors declare that no competing interests exist.
  7. Stuart J Edelstein

    Biologie Cellulaire de la Synapse, Inserm U1024, CNRS 8197, Institute of Biology, Ecole Normale Supérieure, Paris, France
    Competing interests
    The authors declare that no competing interests exist.
  8. Crina M Nimigean

    Department of Anesthesiology, Weill Cornell Medicine, New York, United States
    Competing interests
    The authors declare that no competing interests exist.
  9. Delphine Joseph

    BioCIS, Université Paris-Sud, Châtenay-Malabry, France
    Competing interests
    The authors declare that no competing interests exist.
  10. Pierre-Jean Corringer

    Channel Receptors Unit, Institut Pasteur, Paris, France
    For correspondence
    pjcorrin@pasteur.fr
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-4770-430X

Funding

Agence Nationale de la Recherche (Pentagate)

  • Anaïs Menny
  • Solène N Lefebvre
  • Emmanuelle Drège
  • Zaineb Fourati
  • Marc Delarue
  • Delphine Joseph
  • Pierre-Jean Corringer

Centre National de la Recherche Scientifique (UMR 3571)

  • Anaïs Menny
  • Solène N Lefebvre
  • Delphine Joseph

Institut Pasteur

  • Anaïs Menny
  • Solène N Lefebvre
  • Zaineb Fourati
  • Marc Delarue
  • Pierre-Jean Corringer

Université Pierre et Marie Curie (PhD student fellowship)

  • Anaïs Menny
  • Solène N Lefebvre

Fondation pour la Recherche Médicale (DEQ20140329497)

  • Anaïs Menny
  • Pierre-Jean Corringer

National Institutes of Health (R01 GM088352)

  • Crina M Nimigean

Centre National de la Recherche Scientifique (UMR 8076)

  • Emmanuelle Drège
  • Delphine Joseph

Centre National de la Recherche Scientifique (UMR 3528)

  • Zaineb Fourati
  • Marc Delarue

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

Reviewing Editor

  1. Baron Chanda, University of Wisconsin-Madison, United States

Publication history

  1. Received: December 7, 2016
  2. Accepted: March 14, 2017
  3. Accepted Manuscript published: March 15, 2017 (version 1)
  4. Version of Record published: April 20, 2017 (version 2)
  5. Version of Record updated: May 9, 2017 (version 3)

Copyright

© 2017, Menny 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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Further reading

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