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Structural analysis for vacuolar type ATPase to reveal molecular mechanism of rotary proton translocation

Research Project

Project/Area Number 17H03648
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Structural biochemistry
Research InstitutionKyoto Sangyo University

Principal Investigator

YOKOYAMA Ken  京都産業大学, 生命科学部, 教授 (70271377)

Co-Investigator(Kenkyū-buntansha) 光岡 薫  大阪大学, 超高圧電子顕微鏡センター, 教授 (60301230)
Project Period (FY) 2017-04-01 – 2020-03-31
Project Status Completed (Fiscal Year 2019)
Budget Amount *help
¥17,940,000 (Direct Cost: ¥13,800,000、Indirect Cost: ¥4,140,000)
Fiscal Year 2019: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Fiscal Year 2018: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
Fiscal Year 2017: ¥11,050,000 (Direct Cost: ¥8,500,000、Indirect Cost: ¥2,550,000)
Keywordsクライオ電子顕微鏡 / 構造解析 / 生体エネルギー変換 / V-ATPase / ATP合成 / エネルギー変換 / 分子モーター / 構造生物学 / 原子モデル / ATP合成酵素 / bioenergetics / ATP 合成酵素 / cryoEM / ATP synthase / 単粒子解析 / bioenergatics / rotary motor
Outline of Final Research Achievements

ATP synthase is an energy-converting molecular machine that converts the flow of protons through the membrane domain into rotational motion to synthesize ATP. In this study, the overall structure of V/A-type ATP synthase from thermophilic bacteria was clarified by single-particle analysis using cryo-electron microscopy. In addition, three rotation states were identified, enabled us to know motion of peripheral stalks during rotation. Then, the Vo portion was isolated and a density map of the atomic resolution was obtained by single-particle analysis using cryo electron microscope, indicating proton pathway from both sides of the membrane. Using these structures, we were able to construct an atomic model of the intact ATP synthase.

Academic Significance and Societal Importance of the Research Achievements

クライオ電子顕微鏡による構造解析は、近年急速に発展した分野であり、生命現象を原子レベルで理解することを可能にする強力な手法である。また、タンパク質を標的とした薬剤設計に有効で、たとえばコロナウイルスの膜タンパク質に対する薬剤開発も可能にする。我々は、早い段階でこの手法に着目し、国内でははじめて複雑な膜タンパク質複合体の構造解析に成功し、この分野の進展に大きく貢献する成果となった。

Report

(4 results)
  • 2019 Annual Research Report   Final Research Report ( PDF )
  • 2018 Annual Research Report
  • 2017 Annual Research Report
  • Research Products

    (24 results)

All 2019 2018 2017 Other

All Journal Article (4 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 3 results,  Open Access: 4 results) Presentation (18 results) (of which Int'l Joint Research: 3 results,  Invited: 6 results) Remarks (2 results)

  • [Journal Article] Cryo-EM studies of the rotary H<sup>+</sup>-ATPase/synthase from <i>Thermus thermophilus</i>2019

    • Author(s)
      Nakanishi Atsuko、Kishikawa Jun-ichi、Mitsuoka Kaoru、Yokoyama Ken
    • Journal Title

      Biophysics and Physicobiology

      Volume: 16 Issue: 0 Pages: 140-146

    • DOI

      10.2142/biophysico.16.0_140

    • NAID

      130007701812

    • ISSN
      2189-4779
    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Cryo EM structure of intact rotary H+-ATPase/synthase from Thermus thermophilus.2018

    • Author(s)
      #Nakanishi A, #Kishikawa J (#共同筆頭著者), Tamakoshi M, Mitsuoka K, Yokoyama K.
    • Journal Title

      Nat Commun.

      Volume: 9 Issue: 1 Pages: 89-89

    • DOI

      10.1038/s41467-017-02553-6

    • NAID

      40021660371

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] General anesthetics cause mitochondrial dysfunction and reduction of intracellular ATP levels.2018

    • Author(s)
      Kishikawa JI, Inoue Y, Fujikawa M, Nishimura K, Nakanishi A, Tanabe T, Imamura H, Yokoyama K.
    • Journal Title

      PLoS ONE

      Volume: 13 Issue: 1 Pages: e0190213-e0190213

    • DOI

      10.1371/journal.pone.0190213

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] クライオ電子顕微鏡による液胞型 ATPase (VoV1) の構造解析2017

    • Author(s)
      岸川淳一、中西温子、光岡薫、横山謙
    • Journal Title

      京都産業大学研究所報

      Volume: 10 Pages: 99-101

    • Related Report
      2017 Annual Research Report
    • Open Access
  • [Presentation] 好熱菌 V/A-ATPase のヌクレオチドフリー構造の解析2019

    • Author(s)
      古田綾、岸川淳一、中西温子、光岡薫、横山謙
    • Organizer
      第45回日本生体エネルギー研究会討論会
    • Related Report
      2019 Annual Research Report
  • [Presentation] 好熱菌 V型 ATP合成酵素の膜内在性ドメイン Voの構造解析2019

    • Author(s)
      岸川淳一、中西温子、古田綾、加藤貴之、光岡薫、横山謙
    • Organizer
      第45回日本生体エネルギー研究会討論会
    • Related Report
      2019 Annual Research Report
  • [Presentation] キメラ軸を持つV1-ATPaseの1分子解析2019

    • Author(s)
      三谷奈穂、中野敦樹、岸川淳一、横山謙
    • Organizer
      第45回日本生体エネルギー研究会討論会
    • Related Report
      2019 Annual Research Report
  • [Presentation] Nanodisc に再構成した好熱菌由来 V-ATPase の単粒子解析2019

    • Author(s)
      中西温子、岸川淳一、玉腰雅忠、光岡薫、横山謙
    • Organizer
      生理研研究会 岡崎市
    • Related Report
      2019 Annual Research Report
    • Invited
  • [Presentation] Single-particle analysis of lipid nanodisc-reconstituted V-ATPase from Thermus thermophilus2019

    • Author(s)
      中西温子、岸川淳一、玉腰雅忠、光岡薫、横山謙
    • Organizer
      第57回日本生物物理学会 宮崎市
    • Related Report
      2019 Annual Research Report
  • [Presentation] Single particle analysis of membrane Vo domain of V type ATP synthase2019

    • Author(s)
      岸川淳一、 加藤 貴之、 古田 綾、 中西 温子、 光岡 薫、 横山 謙
    • Organizer
      蛋白質科学会年会
    • Related Report
      2019 Annual Research Report
    • Invited
  • [Presentation] ヒト V-ATPase の発現系構築およびトモグラフィー解析の試み2018

    • Author(s)
      中西温子、岸川淳一、表弘志、光岡薫、横山謙
    • Organizer
      第43回日本生体エネルギー研究会討論会
    • Related Report
      2018 Annual Research Report
  • [Presentation] クライオ電子顕微鏡による好熱菌 Thermus thermophilus 由来 V 型 ATP 合成酵素の単粒子解析2018

    • Author(s)
      中西温子、岸川淳一、玉腰雅忠、光岡薫、横山謙
    • Organizer
      第56回日本生物物理学会
    • Related Report
      2018 Annual Research Report
    • Invited
  • [Presentation] クライオ電子顕微鏡による VoV1 の単粒子解析2018

    • Author(s)
      中西温子、岸川淳一、玉腰雅忠、光岡薫、横山謙
    • Organizer
      第18回日本蛋白質科学会
    • Related Report
      2018 Annual Research Report
  • [Presentation] Structural dynamics of V-ATPase via cryo-EM single molecule analysis2018

    • Author(s)
      岸川 淳一、中西 温子、玉腰 雅忠、光岡 薫、横山 謙
    • Organizer
      第18回日本蛋白質科学会年会
    • Related Report
      2018 Annual Research Report
  • [Presentation] De novo designed axis works as a rotor of rotary motor2018

    • Author(s)
      Jun-ichi Kishikawa, Mihori Baba, Atsuko Nakanishi, Kaoru Mitsuoka, Ken Yokoyama
    • Organizer
      European Bioenergetics Conference 2018
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Cryo-tomography and sub-tomogram averaging of dimeric F type ATP synthase at bovine sub-mitochondrial particle2018

    • Author(s)
      Jun-ichi Kishikawa, Atsuko Nakanishi, Masatoshi Murai, Kaoru Mitsuoka, Ken Yokoyama
    • Organizer
      第56回日本生物物理学会年会
    • Related Report
      2018 Annual Research Report
  • [Presentation] はじめてのクライオ電顕2017

    • Author(s)
      横山 謙
    • Organizer
      第43回生体エネルギー研究会討論会
    • Related Report
      2017 Annual Research Report
    • Invited
  • [Presentation] クライオ EM による好熱菌由来 V 型 ATP 合成酵素の単粒子解析2017

    • Author(s)
      岸川 淳一、中西 温子、光岡 薫、横山 謙
    • Organizer
      ConBio2017
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Single-particle analysis of V-type ATPase/synthase from Thermus thermophirus by cryo-EM2017

    • Author(s)
      中西温子、岸川淳一、光岡薫、横山謙
    • Organizer
      第55回日本生物物理学会
    • Related Report
      2017 Annual Research Report
    • Invited
  • [Presentation] 低温電子顕微鏡を用いたトモグラフィ法によるミトコンドリア膜タンパク質複合体の構造解析の試み2017

    • Author(s)
      岸川 淳一、中西 温子、光岡 薫、横山 謙
    • Organizer
      第43回生体エネルギー研究会討論会
    • Related Report
      2017 Annual Research Report
  • [Presentation] Cryo-EM structure of intact rotary H+-ATPase/synthase from Thermus thermophilus2017

    • Author(s)
      中西温子、岸川淳一、光岡薫、横山謙
    • Organizer
      ConBio2017
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Single-particle analysis of Thermus thermophilus VoV1 by Cryo-EM2017

    • Author(s)
      中西温子、岸川淳一、横山謙
    • Organizer
      第17回日本蛋白質科学会
    • Related Report
      2017 Annual Research Report
  • [Remarks] ようこそ 生体膜エネルギー研究室へ

    • URL

      http://www.cc.kyoto-su.ac.jp/~yokoken/index-j.htm

    • Related Report
      2019 Annual Research Report
  • [Remarks] ようこそ、生体膜エネルギー研究室へ

    • URL

      http://www.cc.kyoto-su.ac.jp/~yokoken/index-j.htm

    • Related Report
      2017 Annual Research Report

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Published: 2017-04-28   Modified: 2021-02-19  

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