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Functional analysis of mitochondrial VDAC using synthetic ubiquinone probes

Research Project

Project/Area Number 15K07411
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Bioorganic chemistry
Research InstitutionKyoto University

Principal Investigator

MURAI MASATOSHI  京都大学, 農学研究科, 助教 (80543925)

Co-Investigator(Renkei-kenkyūsha) YAMAMOTO TAKENORI  徳島大学, 先端酵素学研究所, 講師 (80457324)
Project Period (FY) 2015-04-01 – 2018-03-31
Project Status Completed (Fiscal Year 2017)
Budget Amount *help
¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000)
Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2015: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Keywordsユビキノン / ミトコンドリア / VDAC / 光親和性標識 / ケミカルバイオロジー / 膜透過性亢進 / クリックケミストリー
Outline of Final Research Achievements

The role of the voltage-dependent anion channel (VDAC) as a key link between the mitochondrial matrix and cytosol has been firmly established; however, its involvement in the regulation of mitochondrial permeability transition (PT) remains highly controversial. In the present study, we investigated whether the ubiquinone molecule directly binds to VDAC in Saccharomyces cerevisiae mitochondria by photoaffinity labeling using two photoreactive ubiquinones (PUQ-1 and PUQ-2). We demonstrated that ubiquinone specifically binds to the C-terminal region Phe221-Lys234 of the VDAC, connecting the 15th and 16th β-strand sheets. PUQ-1 and PUQ-2 both significantly suppressed the Ca2+-induced mitochondrial PT (monitored by mitochondrial swelling) at the one digit μM level. The results of the present study provided, for the first time to our knowledge, direct evidence indicating that the ubiquinone molecule specifically binds to VDAC1 through its quinone-head ring.

Report

(4 results)
  • 2017 Annual Research Report   Final Research Report ( PDF )
  • 2016 Research-status Report
  • 2015 Research-status Report
  • Research Products

    (47 results)

All 2018 2017 2016 2015 Other

All Journal Article (12 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 10 results,  Open Access: 1 results,  Acknowledgement Compliant: 7 results) Presentation (30 results) (of which Int'l Joint Research: 4 results,  Invited: 1 results) Remarks (5 results)

  • [Journal Article] Identification of the binding sites for ubiquinone and inhibitors in the Na+-pumping NADH-ubiquinone oxidoreductase from Vibrio cholerae by photoaffinity labeling2017

    • Author(s)
      Ito T., Murai M., Ninokura S., Kitazumi Y., Mezic K.G., Cress B.F., Koffas M.A.G., Morgan J.E., Barquera B., and Miyoshi H.
    • Journal Title

      J. Biol. Chem.

      Volume: 292 Issue: 19 Pages: 7727-7742

    • DOI

      10.1074/jbc.m117.781393

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
  • [Journal Article] Pinpoint Chemical Modification of the Quinone-Access Channel of Mitochondrial Complex I via a Two-Step Conjugation Reaction2017

    • Author(s)
      Masuya Takahiro、Murai Masatoshi、Ito Takeshi、Aburaya Shunsuke、Aoki Wataru、Miyoshi Hideto
    • Journal Title

      Biochemistry

      Volume: 56 Issue: 32 Pages: 4279-4287

    • DOI

      10.1021/acs.biochem.7b00612

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Synthetic Ubiquinones Specifically Bind to Mitochondrial Voltage-Dependent Anion Channel 1 (VDAC1) in Saccharomyces cerevisiae Mitochondria.2017

    • Author(s)
      Masatoshi Murai, Ayaka Okuda, Takenori Yamamoto, Yasuo Shinohara and Hideto Miyoshi
    • Journal Title

      Biochemistry

      Volume: 56 Issue: 4 Pages: 570-581

    • DOI

      10.1021/acs.biochem.6b01011

    • Related Report
      2016 Research-status Report
    • Peer Reviewed
  • [Journal Article] Action mechanism of the inhibitors of mitochondrial respiratory complex I based on structure-activity relationship (SAR) studies2017

    • Author(s)
      三芳秀人,村井正俊
    • Journal Title

      Japanese Journal of Pesticide Science

      Volume: 42 Issue: 1 Pages: 44-48

    • DOI

      10.1584/jpestics.W17-09

    • NAID

      130005995627

    • ISSN
      2187-0365, 2187-8692
    • Related Report
      2016 Research-status Report
  • [Journal Article] Characterization of the ubiquinone-binding protein Coq10 using a synthetic ubiquinone probe2017

    • Author(s)
      村井正俊,三芳秀人
    • Journal Title

      Japanese Journal of Pesticide Science

      Volume: 42 Issue: 1 Pages: 65-70

    • DOI

      10.1584/jpestics.W17-19

    • NAID

      130005995713

    • ISSN
      2187-0365, 2187-8692
    • Related Report
      2016 Research-status Report
  • [Journal Article] Specific methylation of Asp160 (49 kDa subunit) located inside the quinone binding cavity of bovine mitochondrial complex I2016

    • Author(s)
      Murai, M., Inaoka, H., Masuya, T., Aburaya, S., Aoki, W., and Miyoshi, H.
    • Journal Title

      Biochemistry

      Volume: 55 Issue: 23 Pages: 3189-3197

    • DOI

      10.1021/acs.biochem.6b00190

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Characterization of the reaction of decoupling ubiquinone with bovine mitochondrial respiratory complex I2016

    • Author(s)
      Masuya, T., Okuda, K., Murai, M. and Miyoshi, H.
    • Journal Title

      Bioscience, Biotechnology, and Biochemistry

      Volume: 80 Issue: 8 Pages: 1463-1469

    • DOI

      10.1080/09168451.2016.1179095

    • Related Report
      2016 Research-status Report
    • Peer Reviewed
  • [Journal Article] Current topics on inhibitors of respiratory complex I2016

    • Author(s)
      Murai, M. and Miyoshi, H.
    • Journal Title

      Biochim. Biophys. Acta (Bioenergetics)

      Volume: 1857 Issue: 7 Pages: 884-891

    • DOI

      10.1016/j.bbabio.2015.11.009

    • Related Report
      2015 Research-status Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Reduction of synthetic ubiquinone QT catalyzed by bovine mitochondrial complex I is decoupled with proton translocation2016

    • Author(s)
      Okuda, K., Murai, M., Aburaya, S., Aoki, W. and Miyoshi, H.
    • Journal Title

      Biochemistry

      Volume: 55 Issue: 3 Pages: 470-481

    • DOI

      10.1021/acs.biochem.5b01090

    • Related Report
      2015 Research-status Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Murai M, Habu S, Murakami S, Ito T, Miyoshi H2015

    • Author(s)
      Production of new amilorides as potent inhibitors of mitochondrial respiratory complex I
    • Journal Title

      Bioscience, Biotechnology, and Biochemistry

      Volume: in press Issue: 7 Pages: 1061-1066

    • DOI

      10.1080/09168451.2015.1010479

    • Related Report
      2015 Research-status Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Identification of the binding position of amilorides in the quinone binding pocket of mitochondrial complex I2015

    • Author(s)
      Ito, T. Murai, M., Morisaka, H., and Miyoshi, H.
    • Journal Title

      Biochemistry

      Volume: 54 Issue: 23 Pages: 3677-3686

    • DOI

      10.1021/acs.biochem.5b00385

    • Related Report
      2015 Research-status Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Amilorides bind to the quinone binding pocket of bovine miyochondrial complex I2015

    • Author(s)
      Murai, M., Murakami, S., Ito, T., and Miyoshi, H.
    • Journal Title

      Biochemistry

      Volume: in press Issue: 17 Pages: 2739-2746

    • DOI

      10.1021/acs.biochem.5b00187

    • Related Report
      2015 Research-status Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Presentation] 出芽酵母ミトコンドリアのVDACを阻害するペンテンジオール類縁体の作用機構研究2018

    • Author(s)
      運天優歩、村井正俊、一丸直哉、山本武範、篠原康雄、三芳秀人
    • Organizer
      日本農芸化学会2018年度大会
    • Related Report
      2017 Annual Research Report
  • [Presentation] 呼吸鎖複合体-Iにおけるアミロライド類の結合部位の同定2018

    • Author(s)
      木村洋則、村井正俊、三芳秀人
    • Organizer
      日本農芸化学会2018年度大会
    • Related Report
      2017 Annual Research Report
  • [Presentation] ミトコンドリアADP/ATP 輸送体を阻害するエポキシシクロヘキセンジオン類の作用機構研究2018

    • Author(s)
      青山綾希、村井正俊、一丸直哉、三芳秀人
    • Organizer
      日本農芸化学会2018年度大会
    • Related Report
      2017 Annual Research Report
  • [Presentation] コレラ菌Na+輸送型NADH-キノン酸化還元酵素における阻害剤結合部位の同定2018

    • Author(s)
      伊藤剛、村井正俊、二ノ倉聡、北隅優希、Blanca Barquera、三芳秀人
    • Organizer
      日本農芸化学会2018年度大会
    • Related Report
      2017 Annual Research Report
  • [Presentation] 合成ユビキノンは出芽酵母ミトコンドリア電位依存性アニオンチャンネル(VDAC)に特異的に結合する2017

    • Author(s)
      村井正俊、奥田絢香、山本武範、篠原康雄、三芳秀人
    • Organizer
      日本農芸化学会2017年度大会
    • Place of Presentation
      京都女子大学
    • Year and Date
      2017-03-20
    • Related Report
      2016 Research-status Report
  • [Presentation] コレラ菌NADH-キノン酸化還元酵素の特異的阻害剤コロルミシンの単離と全合成2017

    • Author(s)
      伊藤剛、戸崎龍彦、村井正俊、Barquera B, 三芳秀人
    • Organizer
      日本農芸化学会2017年度大会
    • Place of Presentation
      京都女子大学
    • Year and Date
      2017-03-20
    • Related Report
      2016 Research-status Report
  • [Presentation] ミトコンドリア呼吸鎖複合体-Iの位置特異的化学修飾2017

    • Author(s)
      桝谷貴洋、村井正俊、三芳秀人
    • Organizer
      日本農芸化学会2017年度大会
    • Place of Presentation
      京都女子大学
    • Year and Date
      2017-03-20
    • Related Report
      2016 Research-status Report
  • [Presentation] トシル化学によるミトコンドリア複合体-Iのユビキノン結合部位の特異的メチル化2017

    • Author(s)
      桝谷貴洋、稲岡宏幸、村井正俊、三芳秀人
    • Organizer
      日本農薬学会第42回大会
    • Place of Presentation
      愛媛大学
    • Year and Date
      2017-03-08
    • Related Report
      2016 Research-status Report
  • [Presentation] 合成ユビキノンは出芽酵母ミトコンドリア電位依存性アニオンチャンネル(VDAC)に特異的に結合する2017

    • Author(s)
      村井正俊、奥田絢香、山本武範、篠原康雄、三芳秀人
    • Organizer
      日本農薬学会第42回大会
    • Place of Presentation
      愛媛大学
    • Year and Date
      2017-03-08
    • Related Report
      2016 Research-status Report
  • [Presentation] コレラ菌NADH-キノン酸化還元酵素の阻害剤コロルミシンの単離と全合成研究2017

    • Author(s)
      伊藤剛、戸崎龍彦、村井 正俊、Blanca Barquera, 三芳 秀人
    • Organizer
      日本農薬学会第42回大会
    • Place of Presentation
      愛媛大学
    • Year and Date
      2017-03-08
    • Related Report
      2016 Research-status Report
  • [Presentation] ミトコンドリアADP/ATP輸送体を阻害するエポキシシクロヘキセンジオン類の作用機構研究2017

    • Author(s)
      青山綾希、村井正俊、三芳秀人
    • Organizer
      日本農芸化学会 関西・中四国・西日本支部2017年度合同大会
    • Related Report
      2017 Annual Research Report
  • [Presentation] ミトコンドリア呼吸鎖複合体-Iのユビキノン結合ポケットの位置特異的化学修飾2017

    • Author(s)
      村井正俊、桝谷貴洋、三芳秀人
    • Organizer
      日本農芸化学会 関西・中四国・西日本支部2017年度合同大会
    • Related Report
      2017 Annual Research Report
  • [Presentation] コレラ菌Na+輸送型NADH-キノン酸化還元酵素における阻害剤結合部位の同定2017

    • Author(s)
      伊藤剛、村井正俊、Barquera B、三芳秀人
    • Organizer
      日本農芸化学会 関西・中四国・西日本支部 2017年度合同大阪大会
    • Related Report
      2017 Annual Research Report
  • [Presentation] コレラ菌Na+輸送性NADH-キノン酸化還元酵素の阻害剤結合部位の同定2017

    • Author(s)
      伊藤剛、二ノ倉聡、村井正俊、北隅優希、Blanca Barquera、三芳秀人
    • Organizer
      日本生体エネルギー研究会第43回討論会
    • Related Report
      2017 Annual Research Report
  • [Presentation] 呼吸鎖複合体-Iのキノン結合ポケットのピンポイント化学修飾2017

    • Author(s)
      村井正俊、桝谷貴洋、三芳秀人
    • Organizer
      日本生体エネルギー研究会第43回討論会
    • Related Report
      2017 Annual Research Report
  • [Presentation] Characterization of the ubiquinone-binding pocket of respiratory complex I through chemical modifications2017

    • Author(s)
      Masatoshi Murai
    • Organizer
      The 2017 Gordon Research Conference on Bioenergetics
    • Related Report
      2017 Annual Research Report
  • [Presentation] ミトコンドリア呼吸鎖複合体-Iにおける合成ユビキノンQTの還元はプロトン輸送と共役しない2016

    • Author(s)
      村井正俊、奥田健司、桝谷貴洋、三芳秀人
    • Organizer
      日本生体エネルギー研究会第42回討論会
    • Place of Presentation
      名古屋工業大学
    • Year and Date
      2016-12-19
    • Related Report
      2016 Research-status Report
  • [Presentation] ミトコンドリア呼吸鎖複合体-Iの位置特異的化学修飾2016

    • Author(s)
      桝谷貴洋、村井正俊、稲岡宏幸、三芳秀人
    • Organizer
      日本生体エネルギー研究会第42回討論会
    • Place of Presentation
      名古屋工業大学
    • Year and Date
      2016-12-19
    • Related Report
      2016 Research-status Report
  • [Presentation] Identification of the binding site of amilorides in mitochondrial respiratory complex I2016

    • Author(s)
      Ito T, Murai M and Miyoshi H
    • Organizer
      18th European Bioenergetics Conference (EBEC2016)
    • Place of Presentation
      Riva del Garda Congressi, Italy
    • Year and Date
      2016-07-02
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research
  • [Presentation] Synthetic ubiquinones specifically bind to mitochondrial voltage-dependent anion channel of Saccharomyces cerevisiae2016

    • Author(s)
      Murai M, Okuda A, Yamamoto T, Shinohara Y and Miyoshi H
    • Organizer
      18th European Bioenergetics Conference (EBEC2016)
    • Place of Presentation
      Riva del Garda Congressi, Italy
    • Year and Date
      2016-07-02
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research
  • [Presentation] Site-specific chemical modifications of the quinone binding cavity of bovine heart mitochondrial complex I2016

    • Author(s)
      Masuya T, Murai M, Inaoka H and Miyoshi H
    • Organizer
      18th European Bioenergetics Conference (EBEC2016)
    • Place of Presentation
      Riva del Garda Congressi, Italy
    • Year and Date
      2016-07-02
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research
  • [Presentation] ミトコンドリア複合体-Iによる合成ユビキノンQTの還元はプロトン輸送と共役しない2016

    • Author(s)
      奥田健司、村井正俊、油屋駿介、青木航、三芳秀人
    • Organizer
      日本農芸化学会2016年度大会
    • Place of Presentation
      札幌コンベンションセンター
    • Year and Date
      2016-03-27
    • Related Report
      2015 Research-status Report
  • [Presentation] トシル化学によるミトコンドリア複合体-Iのユビキノン結合部位の特異的メチル化2016

    • Author(s)
      稲岡宏幸、村井正俊、桝谷貴洋、油屋駿介、青木航、三芳秀人
    • Organizer
      日本農芸化学会2016年度大会
    • Place of Presentation
      札幌コンベンションセンター
    • Year and Date
      2016-03-27
    • Related Report
      2015 Research-status Report
  • [Presentation] 出芽酵母ミトコンドリアVDACはユビキノンと特異的に結合する2016

    • Author(s)
      奥田絢香、村井正俊、山本武範、篠原康雄、三芳秀人
    • Organizer
      日本農芸化学会2016年度大会
    • Place of Presentation
      札幌コンベンションセンター
    • Year and Date
      2016-03-27
    • Related Report
      2015 Research-status Report
  • [Presentation] ミトコンドリア複合体-I阻害剤としてのアミロライド類の作用機構研究2016

    • Author(s)
      伊藤剛、村井正俊、村上園実、森坂裕信、三芳秀人
    • Organizer
      日本農薬学会第41回大会
    • Place of Presentation
      島根大学
    • Year and Date
      2016-03-17
    • Related Report
      2015 Research-status Report
  • [Presentation] ミトコンドリア複合体-Iによる合成ユビキノンQTの還元はプロトン輸送と共役しない2016

    • Author(s)
      桝谷貴洋,奥田健司,村井正俊,三芳秀人
    • Organizer
      日本農薬学会第41回大会
    • Place of Presentation
      島根大学
    • Year and Date
      2016-03-17
    • Related Report
      2015 Research-status Report
  • [Presentation] ミトコンドリア呼吸鎖活性を制御するユビキノン結合性タンパク質Coq10の機能2016

    • Author(s)
      村井正俊、三芳秀人
    • Organizer
      日本農薬学会第41回大会(シンポジウム)
    • Place of Presentation
      島根大学
    • Year and Date
      2016-03-17
    • Related Report
      2015 Research-status Report
    • Invited
  • [Presentation] アミロライド類のミトコンドリア複合体-Iにおける結合部位の同定2015

    • Author(s)
      伊藤剛、村井正俊、村上園実、森坂裕信、三芳秀人
    • Organizer
      第30回農薬デザイン研究会
    • Place of Presentation
      メルパルク京都
    • Year and Date
      2015-11-12
    • Related Report
      2015 Research-status Report
  • [Presentation] ミトコンドリア複合体-Iのピンポイント化学修飾2015

    • Author(s)
      桝谷貴洋、村井正俊、森坂裕信、三芳秀人
    • Organizer
      第30回農薬デザイン研究会
    • Place of Presentation
      メルパルク京都
    • Year and Date
      2015-11-12
    • Related Report
      2015 Research-status Report
  • [Presentation] Amilorides bind to the quinone binding pocket of bovine mitochondrial complex I2015

    • Author(s)
      Murai M, Miyoshi H
    • Organizer
      Gordon Research Conference on Bioenergetics
    • Place of Presentation
      Proctor Academy (USA)
    • Year and Date
      2015-06-21
    • Related Report
      2015 Research-status Report
    • Int'l Joint Research
  • [Remarks] 所属研究室ホームページ

    • URL

      http://www.biofunc-chem.kais.kyoto-u.ac.jp

    • Related Report
      2017 Annual Research Report 2016 Research-status Report 2015 Research-status Report
  • [Remarks] Google Scholar

    • URL

      https://scholar.google.co.jp/citations?user=I66ybkoAAAAJ&hl=ja

    • Related Report
      2017 Annual Research Report 2015 Research-status Report
  • [Remarks] Researchmap

    • URL

      https://researchmap.jp/read0145924/?lang=japanese

    • Related Report
      2017 Annual Research Report
  • [Remarks] Google Scholar

    • URL

      https://scholar.google.com/citations?user=I66ybkoAAAAJ&hl=ja

    • Related Report
      2016 Research-status Report
  • [Remarks] Research map

    • URL

      http://researchmap.jp/read0145924/?lang=japanese

    • Related Report
      2015 Research-status Report

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Published: 2015-04-16   Modified: 2019-03-29  

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