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A new proteomics sample processing method of intracellular insoluble aggregates

Research Project

Project/Area Number 15K14427
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Genome biology
Research Institution防衛大学校(総合教育学群、人文社会科学群、応用科学群、電気情報学群及びシステム工学群)

Principal Investigator

Ichimura Tohru  防衛大学校(総合教育学群、人文社会科学群、応用科学群、電気情報学群及びシステム工学群), 応用科学群, 教授 (50213012)

Co-Investigator(Kenkyū-buntansha) 竹清 貴浩  防衛大学校(総合教育学群、人文社会科学群、応用科学群、電気情報学群及びシステム工学群), 応用科学群, 准教授 (00545981)
八谷 如美  地方独立行政法人東京都立産業技術研究センター, 開発本部開発第二部バイオ応用技術グループ, 主任研究員 (30408075)
田岡 万悟  首都大学東京, 理工学研究科, 准教授 (60271160)
Project Period (FY) 2015-04-01 – 2018-03-31
Project Status Completed (Fiscal Year 2017)
Budget Amount *help
¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2015: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Keywordsプロテオミクス / アグリゲート / LC-MS / BOPs / トリプシン消化 / 質量分析 / 固相抽出 / レーザーカッター / TRIM32
Outline of Final Research Achievements

Protein aggregation causes severe damages to cell and tissue functions. In the present study, we developed a new proteomics sample preparation method that is applicable to minute amounts of aggregates including those isolated from cultured cell lines with laser microdissection systems. This method, which we call BOPs, combines two advantages of current techniques, (1) unbiased binding of reversed-phase polymeric microbeads to any type of protein and (2) enhanced trypsin digestion efficiency in CH3CN-aqueous solvent systems, into a single-tube workflow. BOPs can readily be adapted to a wide range of diluted or denaturant-contaminated samples with high sensitivity, reproducibility, and reliability. The availability of the present BOPs make it especially attractive for next-stage proteomics of rare and limited aggregates.

Report

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

    (7 results)

All 2017 2015

All Journal Article (2 results) (of which Peer Reviewed: 2 results,  Open Access: 1 results,  Acknowledgement Compliant: 1 results) Presentation (5 results)

  • [Journal Article] A Sensitive Microbead-Based Organic Media-Assisted Method for Proteomics Sample Preparation from Dilute and Denaturing Solutions2017

    • Author(s)
      Taoka Masato、Fujii Michihiko、Tsuchiya Masahiro、Uekita Takamasa、Ichimura Tohru
    • Journal Title

      ACS Applied Materials & Interfaces

      Volume: 9 Issue: 49 Pages: 42661-42667

    • DOI

      10.1021/acsami.7b16095

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed
  • [Journal Article] TRIM32-Cytoplasmic-Body Formation Is an ATP-Consuming Process Stimulated by HSP70 in Cells2017

    • Author(s)
      Kawaguchi Y, Taoka M, Takekiyo T, Uekita T, Shoji I, Hachiya N, Ichimura T
    • Journal Title

      PLOS ONE

      Volume: -

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Presentation] An improved on-bead proteolysis procedure for micro-proteomics with denaturating solvent systems2017

    • Author(s)
      Ichimura T, Sakuma T
    • Organizer
      第40回日本基礎老化学会大会
    • Related Report
      2017 Annual Research Report
  • [Presentation] 変性剤混入サンプルの超高感度プロテオミクス分析2017

    • Author(s)
      市村 徹,田岡万梧
    • Organizer
      第90回日本生化学会大会
    • Related Report
      2017 Annual Research Report
  • [Presentation] HSP70によるTRIM32封入体の形成促進2017

    • Author(s)
      市村 徹、河口祐樹、八谷如美
    • Organizer
      平成28年度日本生化学会関東支部例会
    • Place of Presentation
      慈恵医大
    • Related Report
      2016 Research-status Report
  • [Presentation] 分子シャペロンHSP70はアグリゲート形成を促進する2017

    • Author(s)
      市村 徹、河口祐樹、八谷如美
    • Organizer
      第89回日本生化学会大会
    • Place of Presentation
      仙台国際センター
    • Related Report
      2016 Research-status Report
  • [Presentation] HSP70のATPase活性はTRIM32のcytoplasmic body形成に必要である2015

    • Author(s)
      市村徹、河口祐樹、八谷如美
    • Organizer
      BMB2015
    • Place of Presentation
      神戸ポートアイランド
    • Year and Date
      2015-12-01
    • Related Report
      2015 Research-status Report

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Published: 2015-04-16   Modified: 2019-12-27  

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