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Protein-ligand complex formation revealed by fluorescence lifetime-correlation analysis

Research Project

Project/Area Number 15K17814
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeMulti-year Fund
Research Field Physical chemistry
Research InstitutionSaitama University

Principal Investigator

OTOSU Takuhiro  埼玉大学, 研究機構研究企画推進室, 助教 (90564948)

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: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2015: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Keywords蛍光寿命相関分光 / 蛋白質 / 構造ダイナミクス / 蛋白質の構造ダイナミクス / 蛍光相関分光法 / 蛋白質折れ畳み
Outline of Final Research Achievements

Conformational fluctuations of proteins are highly relevant to their biological functions. To this aim, we developed two-dimensional fluorescence lifetime correlation spectroscopy (2D FLCS) which analyze the conformational dynamics of biomolecules in an equilibrium by the correlation analysis of fluorescence lifetime. The aim of this study is to extend 2D FLCS to analyze the conformational dynamics in a non-equilibrium condition. For this purpose, I developed new 2D FLCS instrument which utilizes two laser pulses (UV pulse and visible pulse). UV pulse is used for uncaging the caged compound to transiently increase the ligand concentration in the focal region. The conformational change of protein caused by the interaction with the ligand is then monitored through visible pulse. Preliminary results clearly showed that the instrument developed here is applicable to analyze the ligand-induced conformational changes of proteins with 2D FLCS.

Report

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

    (5 results)

All 2016 2015

All Journal Article (1 results) (of which Peer Reviewed: 1 results,  Open Access: 1 results) Presentation (4 results)

  • [Journal Article] Microsecond protein dynamics observed at the single-molecule level2015

    • Author(s)
      Takuhiro Otosu, Kunihiko Ishii, and Tahei Tahara
    • Journal Title

      Nature communications

      Volume: 6 Issue: 1 Pages: 7685-7685

    • DOI

      10.1038/ncomms8685

    • Related Report
      2015 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] PEG 脂質を導入した支持脂質二重膜の拡散特性2016

    • Author(s)
      猿田萌子,乙須拓洋,山口祥一
    • Organizer
      第54回日本生物物理学会年会
    • Place of Presentation
      つくば国際会議場(茨城県つくば市)
    • Year and Date
      2016-11-25
    • Related Report
      2016 Annual Research Report
  • [Presentation] 新規蛍光相関分光法に基づく生体膜分子の拡散計測に向けた観測サイズ補正法の開発2016

    • Author(s)
      乙須拓洋,山口祥一
    • Organizer
      第10回分子科学討論会
    • Place of Presentation
      神戸ファッションマート(兵庫県神戸市)
    • Year and Date
      2016-09-13
    • Related Report
      2016 Annual Research Report
  • [Presentation] Standing evanescent-wave fluorescence correlation spectroscopy for analyzing the translational diffusion in biomembranes2016

    • Author(s)
      乙須拓洋,山口祥一
    • Organizer
      252nd American Chemical Society National Meeting & Exhibition
    • Place of Presentation
      Pennsylvania Convention Center 「ペンシルべニア州(アメリカ)」
    • Year and Date
      2016-08-21
    • Related Report
      2016 Annual Research Report
  • [Presentation] 定在エバネッセント波を用いた蛍光相関分光装置の開発と脂質二重膜への応用2016

    • Author(s)
      乙須拓洋,山口祥一
    • 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(京都府 京田辺市)
    • Year and Date
      2016-03-26
    • Related Report
      2015 Research-status Report

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

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