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Development of novel protein chemical modification method for imaging analysis of endogenous protein-protein interaction in live cells

Research Project

Project/Area Number 15K17884
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeMulti-year Fund
Research Field Bio-related chemistry
Research InstitutionKyoto University

Principal Investigator

Tamura Tomonori  京都大学, 工学研究科, 助教 (10746639)

Project Period (FY) 2015-04-01 – 2017-03-31
Project Status Completed (Fiscal Year 2016)
Budget Amount *help
¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2016: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2015: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
Keywordsタンパク質化学修飾 / タンパク質ラベリング / 蛋白質化学修飾 / 細胞内有機化学 / 蛍光イメージング / FKBP12
Outline of Final Research Achievements

In this study, we developed a novel chemical method to selectively label proteins expressed endogenously in live cells. Specifically, ligand-directed labeling reagents consisting of "N-acyl-N-alkylsulfonamide (NASA)" as a reactive group were developed. The NASA type labeling reagents were able to covalently modify the target protein more rapidly and efficiently than the conventional ligand directed chemistry. In addition, we successfully demonstrated that this method can selectively modify intracellular endogenous proteins, such as FKBP 12, eDHFR and Hsp90, with high efficiency.

Report

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

    (12 results)

All 2017 2016 2015

All Journal Article (2 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 2 results,  Acknowledgement Compliant: 2 results) Presentation (10 results) (of which Int'l Joint Research: 1 results)

  • [Journal Article] Nucleus-selective Chemical Proteomics Using Hoechst-tagged Reactive Molecules2016

    • Author(s)
      Yuki Yasueda, Tomonori Tamura, Itaru Hamachi
    • Journal Title

      Chem. Lett.

      Volume: 45 Issue: 3 Pages: 265-267

    • DOI

      10.1246/cl.151083

    • NAID

      130005132186

    • Related Report
      2015 Research-status Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Biomembrane-embedded Catalysts for Membrane-associated Protein Labeling on Red Blood Cells2015

    • Author(s)
      Yuki Yasueda, Tomonori Tamura, Keiko Kuwata, Yousuke Takaoka, Itaru Hamachi
    • Journal Title

      Chem. Lett.

      Volume: 44 Issue: 12 Pages: 1673-1675

    • DOI

      10.1246/cl.150797

    • NAID

      130005112533

    • Related Report
      2015 Research-status Report
    • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
  • [Presentation] リガンド指向性NASA化学 (1) : 速度論解析と生細胞タンパク質ラベリング2017

    • Author(s)
      上田 毅、月館 拓、後藤 大輝、田村 朋則、浜地 格
    • Organizer
      日本化学会第97春季年会
    • Place of Presentation
      慶應義塾大学・日吉キャンパス
    • Year and Date
      2017-03-16
    • Related Report
      2016 Annual Research Report
  • [Presentation] リガンド指向性NASA化学 (2) : リガンド親和性と反応性の相関2017

    • Author(s)
      後藤 大輝、月館 拓、上田 毅、田村 朋則、浜地 格
    • Organizer
      日本化学会第97春季年会
    • Place of Presentation
      慶應義塾大学・日吉キャンパス
    • Year and Date
      2017-03-16
    • Related Report
      2016 Annual Research Report
  • [Presentation] リガンド指向性化学によるタンパク質ラベリング(1):LDSP化学における反応性とラベル化効率の定量的評価2016

    • Author(s)
      増田 真理恵、西川 雄貴、鹿又 喬平、松尾 和哉、田村 朋則、浜地 格
    • Organizer
      日本化学会第96春季年会
    • Place of Presentation
      同志社大学・京田辺キャンパス
    • Year and Date
      2016-03-24
    • Related Report
      2015 Research-status Report
  • [Presentation] オルガネラ選択的ケミカルプロテオミクス(2):細胞核タンパク質のプロファイリング2016

    • Author(s)
      藤沢 有磨、安枝 裕貴、田村 朋則、清中 茂樹、浜地 格
    • Organizer
      日本化学会第96春季年会
    • Place of Presentation
      同志社大学・京田辺キャンパス
    • Year and Date
      2016-03-24
    • Related Report
      2015 Research-status Report
  • [Presentation] リガンド指向性化学によるタンパク質ラベリング(2):LDSP化学による内在性タンパク質のFRETセンサー化2016

    • Author(s)
      西川 雄貴、増田 真理恵、松尾 和哉、田村 朋則、浜地 格
    • Organizer
      日本化学会第96春季年会
    • Place of Presentation
      同志社大学・京田辺キャンパス
    • Year and Date
      2016-03-24
    • Related Report
      2015 Research-status Report
  • [Presentation] リガンド指向性化学によるタンパク質ラベリング(5):アシル転移反応を促進する二官能性触媒2016

    • Author(s)
      月館 拓、李 伸、山浦 圭、田村 朋則、浜地 格
    • Organizer
      日本化学会第96春季年会
    • Place of Presentation
      同志社大学・京田辺キャンパス
    • Year and Date
      2016-03-24
    • Related Report
      2015 Research-status Report
  • [Presentation] リガンド指向性化学によるタンパク質ラべリング(4):N‐アシル‐N‐アルキルスルホンアミド化学によるタンパク質ラベル2016

    • Author(s)
      月館 拓、田村 朋則、浜地 格
    • Organizer
      日本化学会第96春季年会
    • Place of Presentation
      同志社大学・京田辺キャンパス
    • Year and Date
      2016-03-24
    • Related Report
      2015 Research-status Report
  • [Presentation] Organelle-selective chemical proteomics(1):Mitochondrial proteome analysis with mitochondria-localizable reactive molecules2016

    • Author(s)
      Tomonori Tamura, Yuki Yasueda, Shigeki Kiyonaka, Itaru Hamachi
    • Organizer
      日本化学会第96春季年会
    • Place of Presentation
      同志社大学・京田辺キャンパス
    • Year and Date
      2016-03-24
    • Related Report
      2015 Research-status Report
  • [Presentation] Organelle-localizable Reactive Molecules : A Chemical Approach for Organellar proteomics in Live Cells2015

    • Author(s)
      Yuki Yasueda, Tomonori Tamura, Shigeki Kiyonaka, Itaru Hamachi
    • Organizer
      IKCOC-13
    • Place of Presentation
      Rihga Royal Hotel KYOTO, JAPAN
    • Year and Date
      2015-09-11
    • Related Report
      2015 Research-status Report
    • Int'l Joint Research
  • [Presentation] リガンド指向型N-acyl-N-alkyl-sulfonamideによる細胞内蛋白質の高効率ラベリング2015

    • Author(s)
      田村 朋則、月館 拓、李 伸、浜地 格
    • Organizer
      第9回バイオ関連化学シンポジウム
    • Place of Presentation
      熊本大学・黒髪南地区キャンパス
    • Year and Date
      2015-09-10
    • Related Report
      2015 Research-status Report

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Published: 2015-04-16   Modified: 2018-03-22  

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