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Development of ultrafast detection system for the single molecule fluorescence based on hybrid photo detector

Research Project

Project/Area Number 26650045
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Biophysics
Research InstitutionTohoku University

Principal Investigator

Takahashi Satoshi  東北大学, 多元物質科学研究所, 教授 (30283641)

Co-Investigator(Renkei-kenkyūsha) KAMAGATA KIYOTO  東北大学, 多元物質科学研究所, 助教 (90432492)
Project Period (FY) 2014-04-01 – 2016-03-31
Project Status Completed (Fiscal Year 2015)
Budget Amount *help
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2015: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000)
Fiscal Year 2014: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Keywords一分子蛍光分光法 / タンパク質 / フォールディング / ライン共焦点顕微鏡 / ライン型共焦点顕微鏡 / 一分子蛍光観察 / ハイブリッド光検出器 / ラインフォーカス型共焦点顕微鏡 / ハイブリッド型フォトダイオード / 石英製顕微対物レンズ
Outline of Final Research Achievements

Single molecule fluorescence spectroscopy (SMFS) is a powerful method for the characterization of protein dynamics; however, the past methods of SMFS has the limit in the time resolution of the fluorescence detection and can not resolve events occurring within 1 ms. In this investigation, we used line confocal microscopy and combined it with the new photodetector, hybrid photodetector (HPD), and enabled to detect fluorescence signals from single molecules at the time resolution of 10 microsecond and for the duration of 10 millisecond. We further characterized the dynamics of a protein, BdpA, and observed that the unfolded BdpA shows various dynamics that should be important for the understanding of protein folding. The developed method can be applied to various proteins and their dynamics at the single molecule level.

Report

(3 results)
  • 2015 Annual Research Report   Final Research Report ( PDF )
  • 2014 Research-status Report
  • Research Products

    (19 results)

All 2015 2014

All Journal Article (5 results) (of which Peer Reviewed: 5 results,  Acknowledgement Compliant: 3 results,  Open Access: 2 results) Presentation (14 results) (of which Invited: 4 results)

  • [Journal Article] Highly Collapsed Conformation of the Initial Folding Intermediates of β-Lactoglobulin with Non-Native α-Helix2015

    • Author(s)
      Tsuyoshi Konuma, Kazumasa Sakurai, Masanori Yagi, Yuji Goto, Teturo Fujisawa, Satoshi Takahashi
    • Journal Title

      Journal of Molecular Biology

      Volume: 427 Issue: 19 Pages: 3158-3165

    • DOI

      10.1016/j.jmb.2015.07.018

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] One-dimensional sliding of p53 along DNA is accelerated in the presence of Ca(2+) or Mg(2+) at millimolar concentrations2015

    • Author(s)
      Murata A, Ito Y, Kashima R, Kanbayashi S, Nanatani K, Igarashi C, Okumura M, Inaba K, Tokino T, Takahashi S, Kamagata K
    • Journal Title

      JOURNAL OF MOLECULAR BIOLOGY

      Volume: 427 Issue: 16 Pages: 2663-2678

    • DOI

      10.1016/j.jmb.2015.06.016

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Complexity of the folding transition of the B domain of protein A revealed by the high-speed tracking of single-molecule fluorescence time series2015

    • Author(s)
      Hiroyuki Oikawa, Kiyoto Kamagata, Munehito Arai, Satoshi Takahashi
    • Journal Title

      J. Phys. Chem. B

      Volume: 119 Issue: 20 Pages: 6081-6091

    • DOI

      10.1021/acs.jpcb.5b00414

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Where the complex things are: single molecule andensemble spectroscopic investigations of protein folding dynamics2015

    • Author(s)
      Satoshi Tkahahshi, Kiyoto Kamagata, Hiroyuki Oikawa
    • Journal Title

      Current Opinion in Structural Biology

      Volume: 36 Pages: 1-9

    • DOI

      10.1016/j.sbi.2015.11.006

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Microsecond Dynamics of a Protein by Single-Molecule Fluorescence Measurements2014

    • Author(s)
      小井川浩之、齊藤雅嵩、高橋聡
    • Journal Title

      Seibutsu Butsuri

      Volume: 54 Issue: 5 Pages: 276-279

    • DOI

      10.2142/biophys.54.276

    • NAID

      130004693958

    • ISSN
      0582-4052, 1347-4219
    • Related Report
      2014 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] ユビキチンの折り畳みダイナミクスの一分子蛍光分光測定2015

    • Author(s)
      齊藤雅嵩, Chen Eric H.-L., Chen Po-Ting, Chen Rita P.-Y., 鎌形清人, 小井川浩之, 高橋聡
    • Organizer
      第9回分子科学討論会
    • Place of Presentation
      東京工業大学(東京都大田区)
    • Year and Date
      2015-09-16
    • Related Report
      2015 Annual Research Report
  • [Presentation] マイクロ秒分解一分子FRET測定よるタンパク質折り畳みダイナミクスの追跡2015

    • Author(s)
      小井川浩之, 新井宗仁, 深澤宏仁, 横田浩章, 井出徹, 高橋聡
    • Organizer
      第9回分子科学討論会
    • Place of Presentation
      東京工業大学(東京都大田区)
    • Year and Date
      2015-09-16
    • Related Report
      2015 Annual Research Report
  • [Presentation] 一分子蛍光分光法によって観測されたユビキチンの変性状態の不均一性2015

    • Author(s)
      齊藤雅嵩、Chen Eric H.-L., Chen Po-Ting, Chen Rita P.-Y., 鎌形清人, 小井川浩之, 高橋聡
    • Organizer
      第53回日本生物物理学会年会
    • Place of Presentation
      金沢大学(石川県金沢市)
    • Year and Date
      2015-09-13
    • Related Report
      2015 Annual Research Report
  • [Presentation] Tracking microsecond single-molecule FRET dynamics on the fast protein folding by the line-confocal microscopy2015

    • Author(s)
      Hiroyuki Oikawa, Munehito Arai, Atsuhito Fukasawa, Hiroaki Yokota , Toru Ide , Satoshi Takahashi
    • Organizer
      第53回日本生物物理学会年会
    • Place of Presentation
      金沢大学(石川県金沢市)
    • Year and Date
      2015-09-13
    • Related Report
      2015 Annual Research Report
  • [Presentation] 一分子蛍光分光法によるタンパク質のフォールディング研究2015

    • Author(s)
      高橋聡
    • Organizer
      機能物性融合科学研究会シリーズ2ソフトダイナミクス
    • Place of Presentation
      東京大学物性研究所(千葉県柏市)
    • Year and Date
      2015-04-02
    • Related Report
      2015 Annual Research Report
    • Invited
  • [Presentation] Development of The Line Confocal System for The Single Molecule Tracking of Folding Dynamics of Proteins2015

    • Author(s)
      Hiroyuki Oikawa , Kiyoto Kamagata, Munehito Arai, Atuhito Fukasawa, Hiroaki Yokota, Toru Ide, Satoshi Takahashi
    • Organizer
      Biophysical Society 59th Annual Meeting
    • Place of Presentation
      Baltimore, USA
    • Year and Date
      2015-02-07 – 2015-02-11
    • Related Report
      2014 Research-status Report
  • [Presentation] Dynamics of Protein Folding Studied by Single Molecule Fluorescence Time Series Measurements at Microsecond Resolution2014

    • Author(s)
      Satoshi Takahashi
    • Organizer
      Indo-Japan Joint Workshop on "Frontiers in Molecular Spectroscopy: Fundamentals and Applications to Material and Biology"
    • Place of Presentation
      Nara
    • Year and Date
      2014-11-25 – 2014-11-28
    • Related Report
      2014 Research-status Report
    • Invited
  • [Presentation] Development of the line confocal system for the single molecule tracking of fast folding dynamics of proteins2014

    • Author(s)
      Hiroyuki Oikawa Kiyoto Kamagata, Munehito Arai, Atsuhito Fukasawa, Hiroaki Yokota, Toru Ide, Satoshi Takahashi
    • Organizer
      第52回日本生物物理学会年会
    • Place of Presentation
      北海道札幌市
    • Year and Date
      2014-09-25 – 2014-09-27
    • Related Report
      2014 Research-status Report
  • [Presentation] Folding dynamics of ubiquitin after rapid mixing detected by single molecule fluorescence spectroscopy2014

    • Author(s)
      Masataka Saito, Chen Erick, Chen Po-Ting, Chen Rita P.-Y., Kiyoto Kamagata, Hiroyuki Oikawa, Satoshi Takahashi
    • Organizer
      第52回日本生物物理学会年会
    • Place of Presentation
      北海道札幌市
    • Year and Date
      2014-09-25 – 2014-09-27
    • Related Report
      2014 Research-status Report
  • [Presentation] Continuous tracking of protein folding at microsecond resolution by a line confocal detection of single molecule fluorescence2014

    • Author(s)
      Satoshi Takahashi
    • Organizer
      The 28th Annual Symposium of The Protein Society
    • Place of Presentation
      San Diego, USA
    • Year and Date
      2014-07-27 – 2014-07-30
    • Related Report
      2014 Research-status Report
    • Invited
  • [Presentation] Development of the line confocal system for the single-molecule tracking of the protein folding transition2014

    • Author(s)
      Hiroyuki Oikawa
    • Organizer
      International Workshop "Over the Barriers of Transition Paths: Dynamical Processes in Proteins and Complex Molecular Systems"
    • Place of Presentation
      Yokohama, Japan
    • Year and Date
      2014-06-28
    • Related Report
      2014 Research-status Report
    • Invited
  • [Presentation] 一分子蛍光分光法によるユビキチンの折り畳みダイナミクス2014

    • Author(s)
      齊藤雅嵩・Chen Hsin-Liang・Chen Po-Ting・Chen Rita P.-Y.・鎌形清人・小井川浩之・高橋聡
    • Organizer
      第14回日本蛋白質科学会年会
    • Place of Presentation
      神奈川県横浜市
    • Year and Date
      2014-06-25 – 2014-06-27
    • Related Report
      2014 Research-status Report
  • [Presentation] 改良したマイクロ秒分解一分子蛍光測定法による蛋白質の高速折り畳みダイナミクスの追跡2014

    • Author(s)
      小井川浩之・鎌形清人・ 新井宗仁・ 深澤宏仁・横田浩章・井出徹・高橋聡
    • Organizer
      第14回日本蛋白質科学会年会
    • Place of Presentation
      神奈川県横浜市
    • Year and Date
      2014-06-25 – 2014-06-27
    • Related Report
      2014 Research-status Report
  • [Presentation] Microsecond-dynamics of the unfolded BdpA and ubiquitin by a line confocal detection of single molecule fluorescence2014

    • Author(s)
      Hiroyuki Oikawa , Masataka Saito, Po-Ting Chen, Hsin-Liang Chen, Rita P.-Y. Chen, Munehito Arai, Kiyoto Kamagata, Satoshi Takahashi
    • Organizer
      The 4th Asia Pacific Protein Association Conference
    • Place of Presentation
      Jeju, Korea
    • Year and Date
      2014-06-17 – 2014-06-20
    • Related Report
      2014 Research-status Report

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Published: 2014-04-04   Modified: 2017-05-10  

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