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In vitro reconstitution of endosomal/lysosomal membrane fusion machinery with purified proteins and defined lipids

Research Project

Project/Area Number 23770117
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeMulti-year Fund
Research Field Structural biochemistry
Research InstitutionOsaka University

Principal Investigator

MIMA Joji  大阪大学, たんぱく質研究所, 准教授 (30335301)

Project Period (FY) 2011 – 2012
Project Status Completed (Fiscal Year 2012)
Budget Amount *help
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2012: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2011: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
Keywords細胞小器官 / 細胞内膜交通 / 細胞内膜融合 / プロテオリポソーム / 試験管内再構成 / SNAREタンパク質 / 蛋白質 / 脂質
Research Abstract

Here, we reported the fusion specificity of reconstituted proteoliposomes bearing purified SNARE proteins in yeast vacuoles and other organelles,to study whether and how SNAREs mediate the compartmental specificity of intracellular membrane fusion. We found that not only vacuolar R-SNARE, but also the non-cognate endosomal and ER-Golgi R-SNAREs caused efficient fusion with vacuolar Qabc-SNAREs. In contrast, their fusion is blocked completely by replacing the vacuolar Qc-SNARE with the non-cognate endosomal counterparts. Our current study establishes that an appropriate assembly of 3Q-SNAREs is crucial for mediating fusion specificity, whereas R-SNARE itself has little contribution to fusion specificity.

Report

(3 results)
  • 2012 Annual Research Report   Final Research Report ( PDF )
  • 2011 Research-status Report
  • Research Products

    (8 results)

All 2013 2012 2011

All Journal Article (2 results) (of which Peer Reviewed: 2 results) Presentation (6 results) (of which Invited: 1 results)

  • [Journal Article] LegC3, an effector protein from Legionella pneumophila, inhibits homotypic yeast vacuole fusion in vivo and in vitro2013

    • Author(s)
      Bennett TL, Kraft SM, Reaves BJ, Mima J, O'Brien KM, Starai VJ
    • Journal Title

      PLoS One

      Volume: 8 Issue: 2 Pages: e56798-e56798

    • DOI

      10.1371/journal.pone.0056798

    • Related Report
      2012 Annual Research Report 2012 Final Research Report
    • Peer Reviewed
  • [Journal Article] Distinct contributions of vacuolar Qabc- and R-SNARE proteins to membrane fusion specificity2012

    • Author(s)
      Izawa R, Onoue T, Furukawa N, Mima J
    • Journal Title

      The Journal of Biological Chemistry

      Volume: 287 Issue: 5 Pages: 3445-3453

    • DOI

      10.1074/jbc.m111.307439

    • Related Report
      2012 Final Research Report 2011 Research-status Report
    • Peer Reviewed
  • [Presentation] 細胞内膜融合マシナリー:再構成SNAREプロテオリポソーム系によるアプローチ.2012

    • Author(s)
      三間穣治
    • Organizer
      第3回神経科学と構造生物学の融合研究会
    • Place of Presentation
      大阪
    • Year and Date
      2012-10-04
    • Related Report
      2012 Final Research Report
  • [Presentation] 細胞内膜融合マシナリー:再構成SNAREプロテオリポソーム系によるアプローチ2012

    • Author(s)
      三間穣治
    • Organizer
      第3回 神経科学と構造生物学の融合研究会
    • Place of Presentation
      大阪大学
    • Related Report
      2012 Annual Research Report
    • Invited
  • [Presentation] Distinct Contributions of Yeast Q- and R-SNAREs to the Specificity of Reconstituted Membrane Fusion.2011

    • Author(s)
      Joji Mima
    • Organizer
      Gordon Research Conference Molecular Membrane Biology
    • Place of Presentation
      Andover, NH, USA
    • Year and Date
      2011-07-13
    • Related Report
      2012 Final Research Report
  • [Presentation] Endolysosomal Membrane Fusion Reconstituted with the Yeast SNAREs, SM Proteins, and Chemically Defined Liposomes.2011

    • Author(s)
      三間穣治
    • Organizer
      第30回内藤コンファレンス「生体膜ダイナミクスと脂質生物学 [II] 」
    • Place of Presentation
      札幌
    • Year and Date
      2011-06-30
    • Related Report
      2012 Final Research Report
  • [Presentation] 酵母液胞オルガネラ膜融合のin vitro完全再構成.2011

    • Author(s)
      三間穣治
    • Organizer
      第11回日本蛋白質科学会年会.
    • Place of Presentation
      大阪
    • Year and Date
      2011-06-07
    • Related Report
      2012 Final Research Report
  • [Presentation] 酵母液胞オルガネラ膜融合のIn vitro完全再構成2011

    • Author(s)
      三間穣治
    • Organizer
      第11回日本蛋白質科学会年会(招待講演)
    • Place of Presentation
      ホテル阪急エキスポパーク
    • Related Report
      2011 Research-status Report

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Published: 2011-08-05   Modified: 2019-07-29  

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