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Investigating protein behaviours in mitochondria by in-organelle NMR

Research Project

Project/Area Number 15K14494
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Biophysics
Research InstitutionTokyo Metropolitan University

Principal Investigator

ITO Yutaka  首都大学東京, 理工学研究科, 教授 (80261147)

Co-Investigator(Kenkyū-buntansha) 美川 務  国立研究開発法人理化学研究所, その他, 研究員 (20321820)
Project Period (FY) 2015-04-01 – 2018-03-31
Project Status Completed (Fiscal Year 2017)
Budget Amount *help
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2017: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2016: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
KeywordsIn-cell NMR / In-organelle NMR / ミトコンドリア / 蛋白質 / in-cell NMR / オルガネラ
Outline of Final Research Achievements

Protein behaviours in eukaryotic organelles were investigated by extending the in-cell NMR approaches to be available for the analysis of proteins localised in organelles, particularly mitochondria.
A model protein with a mitochondrial targeting signal was incorporated into cultured human HeLa.S3 cells by electroporation, and then the mitochondria faction was isolated for NMR experiments. Well-resolved in-mitochondria NMR spectra showed that the proper 3D structure was formed, while different effects of the mitochondria matrix environment to the conformation and dynamics of proteins was suggested from the observed spectral differences when compared with the samples in cytosol or in diluted solution suggested.

Report

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

    (23 results)

All 2018 2017 2016 2015 Other

All Int'l Joint Research (4 results) Journal Article (6 results) (of which Int'l Joint Research: 3 results,  Peer Reviewed: 6 results,  Open Access: 5 results,  Acknowledgement Compliant: 2 results) Presentation (12 results) (of which Int'l Joint Research: 4 results,  Invited: 12 results) Book (1 results)

  • [Int'l Joint Research] University of Cambridge/University of Leicester(United Kingdom)

    • Related Report
      2017 Annual Research Report
  • [Int'l Joint Research] Goethe University Frankfurt(Germany)

    • Related Report
      2017 Annual Research Report
  • [Int'l Joint Research] ETH Zurich(Switzerland)

    • Related Report
      2017 Annual Research Report
  • [Int'l Joint Research] University of Leicester(United Kingdom)

    • Related Report
      2016 Research-status Report
  • [Journal Article] Solution NMR views of dynamical ordering of biomacromolecules2018

    • Author(s)
      Ikeya Teppei、Ban David、Lee Donghan、Ito Yutaka、Kato Koichi、Griesinger Christian
    • Journal Title

      Biochim. Biophys. Acta-General Subjects

      Volume: 1862 Issue: 2 Pages: 287-306

    • DOI

      10.1016/j.bbagen.2017.08.020

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Impact of cellular health conditions on the protein folding state in mammalian cells2017

    • Author(s)
      Inomata Kohsuke、Kamoshida Hajime、Ikari Masaomi、Ito Yutaka、Kigawa Takanori
    • Journal Title

      Chemical Communications

      Volume: 53 Issue: 81 Pages: 11245-11248

    • DOI

      10.1039/c7cc06004a

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Parkinson’s disease-related DJ-1 functions in thiol quality control against aldehyde attack in vitro2017

    • Author(s)
      Matsuda Noriyuki、Kimura Mayumi、Queliconi Bruno Barros、Kojima Waka、Mishima Masaki、Takagi Kenji、Koyano Fumika、Yamano Koji、Mizushima Tsunehiro、Ito Yutaka、Tanaka Keiji
    • Journal Title

      Scientific Reports

      Volume: 7 Issue: 1 Pages: 12816-12816

    • DOI

      10.1038/s41598-017-13146-0

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] In-cell structure determination of proteins at near-physiological concentration2016

    • Author(s)
      Teppei Ikeya, Tomomi Hanashima, Saori Hosoya, Manato Shimazaki, Shiro Ikeda, Masaki Mishima, Peter Guentert and Yutaka Ito
    • Journal Title

      Scientific Reports

      Volume: 6 Issue: 1 Pages: 38312-38312

    • DOI

      10.1038/srep38312

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
  • [Journal Article] A new carbamidemethyl-linked lanthanide chelating tag for PCS NMR spectroscopy of proteins in living HeLa cells.2016

    • Author(s)
      Yuya Hikone, Go Hirai, Masaki Mishima, Kohsuke Inomata, Teppei Ikeya, Souichiro Arai, Masahiro Shirakawa, Mikiko Sodeoka, and Yutaka Ito
    • Journal Title

      Journal of Biomolecular NMR

      Volume: 66 Issue: 2 Pages: 99-110

    • DOI

      10.1007/s10858-016-0059-4

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
  • [Journal Article] In-cell NMRとnonlinear sampling - there is no other way2015

    • Author(s)
      伊藤 隆
    • Journal Title

      分光研究

      Volume: 64 Pages: 413-426

    • NAID

      40020544418

    • Related Report
      2015 Research-status Report
    • Peer Reviewed
  • [Presentation] 常磁性金属を利用したNMRによる生細胞内蛋白質の立体構造解析2018

    • Author(s)
      伊藤 隆
    • Organizer
      日本化学会第98春季年会
    • Related Report
      2017 Annual Research Report
    • Invited
  • [Presentation] NMRによる生細胞内蛋白質の立体構造・ダイナミクスの解析2017

    • Author(s)
      伊藤 隆
    • Organizer
      日本薬学会第137年会
    • Place of Presentation
      仙台国際センター,仙台
    • Year and Date
      2017-03-25
    • Related Report
      2016 Research-status Report
    • Invited
  • [Presentation] in situ structural biology by NMR2017

    • Author(s)
      伊藤 隆
    • Organizer
      分子研研究会「生体金属動態」
    • Related Report
      2017 Annual Research Report
    • Invited
  • [Presentation] NMR approaches to investigate protein folding and dynamics in the crowded intracellular environment2017

    • Author(s)
      伊藤 隆
    • Organizer
      日本生物物理学会第55回年会
    • Related Report
      2017 Annual Research Report
    • Invited
  • [Presentation] Protein structure determination in living eukaryotic cells by in-cell NMR spectroscopy2016

    • Author(s)
      Yutaka Ito
    • Organizer
      第39回日本分子生物学会年会
    • Place of Presentation
      パシフィコ横浜,横浜
    • Year and Date
      2016-12-01
    • Related Report
      2016 Research-status Report
    • Invited
  • [Presentation] In situ structural biology by NMR2016

    • Author(s)
      Yutaka Ito
    • Organizer
      The 42nd Naito Conference “In the Vanguard of Structural Biology: Revolutionizing Life Sciences”
    • Place of Presentation
      シャトレーゼガトーキングダムサッポロ,札幌
    • Year and Date
      2016-10-05
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research / Invited
  • [Presentation] In situ structural biology by NMR2016

    • Author(s)
      Yutaka Ito
    • Organizer
      International Conference on Magnetic Resonance in Biological Systems 2016
    • Place of Presentation
      京都国際会館,京都
    • Year and Date
      2016-08-22
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research / Invited
  • [Presentation] In situ structural biology by NMR2016

    • Author(s)
      Yutaka Ito
    • Organizer
      UK-Japan Symposium “From single molecules to cells and tissues
    • Place of Presentation
      University of Leicester, UK
    • Year and Date
      2016-07-05
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research / Invited
  • [Presentation] NMRを用いた生細胞中の蛋白質の立体構造解析2016

    • Author(s)
      伊藤 隆
    • Organizer
      シンポジウム「細胞環境における蛋白質の動態解析のためのNMRおよび計算科学的アプローチ」
    • Place of Presentation
      京都大学 吉田キャンパス(京都府京都市)
    • Year and Date
      2016-03-28
    • Related Report
      2015 Research-status Report
    • Invited
  • [Presentation] In situ structural biology by NMR2015

    • Author(s)
      Yutaka Ito
    • Organizer
      Pacifichem 2015
    • Place of Presentation
      Honolulu, Hawaii, USA
    • Year and Date
      2015-12-15
    • Related Report
      2015 Research-status Report
    • Int'l Joint Research / Invited
  • [Presentation] In situ structural biology by NMR2015

    • Author(s)
      伊藤 隆
    • Organizer
      BMB2015
    • Place of Presentation
      神戸ポートアイランド(兵庫県神戸市)
    • Year and Date
      2015-12-01
    • Related Report
      2015 Research-status Report
    • Invited
  • [Presentation] 細胞内クラウディング環境における蛋白質のフォールディングとダイナミクスをNMRで観察する2015

    • Author(s)
      伊藤 隆
    • Organizer
      第53回日本生物物理学会年会
    • Place of Presentation
      金沢大学 角間キャンパス(石川県金沢市)
    • Year and Date
      2015-09-14
    • Related Report
      2015 Research-status Report
    • Invited
  • [Book] “Chapter 3, Advances in NMR data acquisition and processing for protein structure determination” Experimental approaches of NMR spectroscopy: Methodology and application to life science and materials science2017

    • Author(s)
      Yutaka Ito and Teppei Ikeya
    • Total Pages
      636
    • Publisher
      Springer
    • Related Report
      2017 Annual Research Report

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

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