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Measurement of proton pump activity by single molecule of VoV1

Research Project

Project/Area Number 26650039
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Functional biochemistry
Research InstitutionKyoto Sangyo University

Principal Investigator

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

Co-Investigator(Renkei-kenkyūsha) YOKOKAWA Ryuji  京都大学, 工学研究科, 准教授 (10411216)
Project Period (FY) 2014-04-01 – 2016-03-31
Project Status Completed (Fiscal Year 2015)
Budget Amount *help
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2014: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
KeywordsV-ATPase / ATP synthase / proton pump / molecular motor / rotation / ATPase / プロトンポンプ / 回転 / 分子モーター
Outline of Final Research Achievements

VoV1 is a rotary molecular motor which pumps protons across the membranes coupled to ATP hydrolysis reaction in V1 portion.
In this project, we first tried to measure proton pump activity of single molecular VoV1 using micro chambers system. The estimated proton/ATP ratio (P/A ratio) is much lower than the theoretical value of 4. This might be due to a leak of proton from the micro chambers. Then we measured the P/A ratio by using a liposome system including pHrode as a pH probe in the membranes. We estimated the P/A ration of 3-4. This result suggests the energy coupling between the V1 and Vo is tight.

Report

(3 results)
  • 2015 Annual Research Report   Final Research Report ( PDF )
  • 2014 Research-status Report
  • Research Products

    (8 results)

All 2015 2014 Other

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

  • [Journal Article] The ingenious structure of central rotor apparatus in VoV1; key for both complex disassembly and energy coupling between V1 and Vo.2015

    • Author(s)
      Atsuko Nakanishi, Jun-ichi Kishikawa, Masatada Tamakoshi, and Ken Yokoyama
    • Journal Title

      PLOS one

      Volume: 10 Issue: 3 Pages: 1-13

    • DOI

      10.1371/journal.pone.0119602

    • Related Report
      2015 Annual Research Report 2014 Research-status Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] GraDeR: membrane protein complex preparation for single particle cryo-EM.2015

    • Author(s)
      Hauer F., Gerle C., Fischer N., Oshima A., Shinzawa-Itoh K., Shimada S., Yokoyama K., Fujiyoshi Y. & Stark H.
    • Journal Title

      Structure

      Volume: 23 Issue: 9 Pages: 1769-1775

    • DOI

      10.1016/j.str.2015.06.029

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Molecular basis of ADP inhibition of vacuolar (V)-type ATPase/synthase.2014

    • Author(s)
      J. Kishikawa, A. Nakanishi, S. Furuike, M. Tamakoshi, K. Yokoyama
    • Journal Title

      J. Biol. Chem.

      Volume: 289 Issue: 1 Pages: 403-412

    • DOI

      10.1074/jbc.m113.523498

    • Related Report
      2014 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] V-ATPase の H+/ATP 比の測定2015

    • Author(s)
      平田かえで、岸川淳一、中西温子、木下一彦、飯田直樹、 横山謙
    • Organizer
      BMB 2015
    • Place of Presentation
      神戸ポートアイランド(神戸市)
    • Year and Date
      2015-12-01
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Single particle analysis of Thermus thermophilus V-ATPase using an electron microscopy ATPase/synthase2015

    • Author(s)
      Atsuko Nakanishi, Nao Takeuchi, Jun-ichi Kishikawa,Kaoru Mistuoka, Ken Yokoyama
    • Organizer
      第53回日本生物物理学会年会
    • Place of Presentation
      金沢大学(石川県 金沢市)
    • Year and Date
      2015-09-13
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Torque generation mechanism in V1 motor2015

    • Author(s)
      BaBa Mihori,Jun-ichi Kishikawa, Atsuko Nakanishi, Nao Takeuchi, Shou Furuike, Ken Yokoyama
    • Organizer
      第53回日本生物物理学会年会
    • Place of Presentation
      金沢大学(石川県 金沢市)
    • Year and Date
      2015-09-13
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Remarks] Ken Yokoyama Lab

    • URL

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

    • Related Report
      2015 Annual Research Report
  • [Remarks] Memebrane BioEnergentics Metabolism Laboratory

    • URL

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

    • Related Report
      2014 Research-status Report

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Published: 2014-04-04   Modified: 2017-05-10  

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