1. Structural Biology and Molecular Biophysics

Mechanical inhibition of isolated Vo from V/A-ATPase for proton conductance

  1. Jun-ichi Kishikawa
  2. Atsuko Nakanishi
  3. Aya Furuta
  4. Takayuki Kato
  5. Keiichi Namba
  6. Masatada Tamakoshi
  7. Kaoru Mitsuoka
  8. Ken Yokoyama  Is a corresponding author
  1. Kyoto Sangyo University, Japan
  2. Osaka University, Japan
  3. Tokyo University of Pharmacy and Life Sciences, Japan
https://doi.org/10.7554/eLife.56862
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Cite this article
  1. Jun-ichi Kishikawa
  2. Atsuko Nakanishi
  3. Aya Furuta
  4. Takayuki Kato
  5. Keiichi Namba
  6. Masatada Tamakoshi
  7. Kaoru Mitsuoka
  8. Ken Yokoyama
(2020)
Mechanical inhibition of isolated Vo from V/A-ATPase for proton conductance
eLife 9:e56862.
https://doi.org/10.7554/eLife.56862
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Abstract

V-ATPase is an energy converting enzyme, coupling ATP hydrolysis/synthesis in the hydrophilic V1 domain, with proton flow through the Vo membrane domain, via rotation of the central rotor complex relative to the surrounding stator apparatus. Upon dissociation from the V1 domain, the Vo domain of the eukaryotic V-ATPase can adopt a physiologically relevant auto-inhibited form in which proton conductance through the Vo domain is prevented, however the molecular mechanism of this inhibition is not fully understood. Using cryo-electron microscopy, we determined the structure of both the holo V/A-ATPase and isolated Vo at near-atomic resolution, respectively. These structures clarify how the isolated Vo domain adopts the auto-inhibited form and how the holo complex prevents formation of the inhibited Vo form.

Data availability

The density maps and the built models for Tth VoV1, Tth V1 (focused refined), and Tth Vo were deposited in EMDB (EMDB ID; 30013, 30014, and 30015) and PDB (PDB ID; 6LY8 for V1 and 6LY9 for isolated Vo), respectively. All data is available in the main text or the supplementary materials.

The following data sets were generated

Article and author information

Author details

  1. Jun-ichi Kishikawa

    Department of Molecular Biosciences, Kyoto Sangyo University, Kyoto, Japan
    Competing interests
    The authors declare that no competing interests exist.

  2. Atsuko Nakanishi

    Department of Molecular Biosciences, Kyoto Sangyo University, Kyoto, Japan
    Competing interests
    The authors declare that no competing interests exist.

  3. Aya Furuta

    Department of Molecular Biosciences, Kyoto Sangyo University, Kyoto, Japan
    Competing interests
    The authors declare that no competing interests exist.

  4. Takayuki Kato

    Institute for Protein Research, Osaka University, Osaka, Japan
    Competing interests
    The authors declare that no competing interests exist.

  5. Keiichi Namba

    Institute for Protein Research, Osaka University, Osaka, Japan
    Competing interests
    The authors declare that no competing interests exist.

  6. Masatada Tamakoshi

    Department of Molecular Biology, Tokyo University of Pharmacy and Life Sciences, Tokyo, Japan
    Competing interests
    The authors declare that no competing interests exist.

  7. Kaoru Mitsuoka

    Research Center for Ultra-High Voltage Electron Microscopy, Osaka University, Osaka, Japan
    Competing interests
    The authors declare that no competing interests exist.

  8. Ken Yokoyama

    Department of Molecular Biosciences, Kyoto Sangyo University, Kyoto, Japan
    For correspondence
    yokoken@cc.kyoto-su.ac.jp
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-6813-1096

Funding

Japan Society for the Promotion of Science (17H03648)

  • Ken Yokoyama

Japan Agency for Medical Research and Development (JP17am0101001)

  • Kaoru Mitsuoka

Ministry of Education, Culture, Sports, Science, and Technology (12024046)

  • Kaoru Mitsuoka

Takeda Science Foundation

  • Ken Yokoyama

Japan Science and Technology Agency (JPMJCR1865)

  • Kaoru Mitsuoka

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

Reviewing Editor

  1. David M Kramer, Michigan State University, United States

Version history

  1. Received: March 12, 2020
  2. Accepted: July 7, 2020
  3. Accepted Manuscript published: July 8, 2020 (version 1)
  4. Version of Record published: July 17, 2020 (version 2)

Copyright

© 2020, Kishikawa 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. Jun-ichi Kishikawa
  2. Atsuko Nakanishi
  3. Aya Furuta
  4. Takayuki Kato
  5. Keiichi Namba
  6. Masatada Tamakoshi
  7. Kaoru Mitsuoka
  8. Ken Yokoyama
(2020)
Mechanical inhibition of isolated Vo from V/A-ATPase for proton conductance
eLife 9:e56862.
https://doi.org/10.7554/eLife.56862

Share this article

https://doi.org/10.7554/eLife.56862

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