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Development of novel probes for single-molecule imaging of proteihs

Research Project

Project/Area Number 19350084
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Chemistry related to living body
Research InstitutionKyoto University

Principal Investigator

YAMAMOTO Yukio  Kyoto University, 高等教育研究開発推進センター, 教授 (90109059)

Co-Investigator(Kenkyū-buntansha) TAKI Masayasu  京都大学, 大学院・地球環境学堂, 助教 (70378850)
Project Period (FY) 2007 – 2008
Project Status Completed (Fiscal Year 2008)
Budget Amount *help
¥19,760,000 (Direct Cost: ¥15,200,000、Indirect Cost: ¥4,560,000)
Fiscal Year 2008: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2007: ¥15,730,000 (Direct Cost: ¥12,100,000、Indirect Cost: ¥3,630,000)
Keywords生体認識・機能化学 / センサー / 希土類蛍光ラベル剤 / エネルギー移動 / タンパク質 / 特異的認識 / 時間分解蛍光 / 一分子計測 / ペプチドタグ / トロポニンC / ジアステレオマー / 蛍光偏光測定 / 二官能基性 / 希土類
Research Abstract

本研究課題では筋収縮弛緩機構の解明を目指し、トロポニンCの-分子計測を可能とする新規なプローブ分子の開発を進めてきた.まず、二官能性蛍光ラペル剤の開発については、二官能基型蛍光ラベル剤BRhoおよびBrosを設計・合成した. HPLCや1H-HMRの解析からBRhoではカルボキシ基の向きにより水素結合様式が変化するため、物性の異なる2つのラベル化タンパク質が生成することがわかった.一方、Brosではタンパク質結合後に一様なコンフォメーションをとることがわかり、蛍光ラベル化剤としてより優れた性質を有することがわかった.次に、タンパク質希土類蛍光ラベル剤の開発を行った.まず、非天然のアミノ酸配列(ペプチドタグ)を特異的に認識する希土類錯体を合成した.本錯体は亜鉛イオンを介してポリアスパラギン酸と錯形成し、ペプチドタグ中のトリプトファン側鎖から希土類中心へのエネルギー移動が効率よく進行することで、希土類の発光が遅延蛍光成分として観測されることがわかった.

Report

(3 results)
  • 2008 Annual Research Report   Final Research Report ( PDF )
  • 2007 Annual Research Report
  • Research Products

    (28 results)

All 2009 2008 2007

All Journal Article (8 results) (of which Peer Reviewed: 8 results) Presentation (20 results)

  • [Journal Article] Selective labeling of tag-fused protein by tryptophan-sensitized luminescence of a terbium complex2009

    • Author(s)
      T. Hirayama, M. Taki, A. Kodan, H. Kato, Y. Yamamoto
    • Journal Title

      Chemical Communications (印刷中)

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] A Rosamine-based Fluorescent Chemosensor for Selective Detection of Silver(I) in Aqueous Solution2008

    • Author(s)
      Shohei Iyoshi, Masayasu Taki, Yukio Yamamoto
    • Journal Title

      Inorganic Chemistry 47

      Pages: 3946-3948

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Effects on Heteroatom Substitution on the Structures, Physicochemical Properties, and Redox Behavior of Nickel(II) Complexes with Pyridine-Containing Macrocyclic2008

    • Author(s)
      Masayasu Taki, Yoshiaki Kawashima, Naoko Sakai, Tasuku Hirayama, Yukio Yamamoto
    • Journal Title

      Ligands, Bulletin of the Chemical Society of Japan 81

      Pages: 590-597

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Colorimetric Response to Mercury -induced Abstraction of Triethylene Glycol Ligand from Gold Nanoparticle Surface2008

    • Author(s)
      Tasuku Hirayama, Masayasu Taki, Yukiyasu Kashiwagi, Masami Nakamoto, Atsushi Kunishita, Shinobu Itoh, Yukio Yamamoto
    • Journal Title

      Dalton Transactions

      Pages: 4705-4707

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Fluorescence Imaging of Intracellular Cadmium Using a Dual-Excitation Ratiometric Chemosensor2008

    • Author(s)
      Masayasu Taki, Mika Desaki, Akio Ojida, Shohei Iyoshi, Tasuku Hirayama, Itaru Hamachi, Yukio Yamamoto
    • Journal Title

      Journal of the American Chemical Society 130

      Pages: 12564-12565

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Development of A Control System for Pulsed-electron Spin Resonance Soectrometers2008

    • Author(s)
      Y. Mizuta, S. Kazama, Y. Ohba, N. Sakai, Y. Yamamoto, Y. Shimoyama
    • Journal Title

      Review of Scientific Instruments 79

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Synthesis of a new bifunctionalised fluorescent label and physical properties of the bound form on model peptide of troponin2007

    • Author(s)
      Tasuku Hirayama, Syohei Iyoshi, Masayasu Taki, Yuichiro Maeda, Yukio Yamamoto
    • Journal Title

      C, Organic & Biomolecular Chemistry 5

      Pages: 2040-2045

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Synthesis of a new bifunctionalised fluorescent label and physical properties of the bound form on model2007

    • Author(s)
      T. Hirayama, S. Iyoshi, M. Taki, Y. Maeda, Y. Yamamoto
    • Journal Title

      Organic & Biomolecular Chemistry 5

      Pages: 2040-2045

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Presentation] ピリジン環含有環状ニッケル錯体と外部配位子との結合挙動2009

    • Author(s)
      旭史悦, 多喜正泰, 山本行男
    • Organizer
      日本化学会第89春季年会
    • Place of Presentation
      日本大学
    • Year and Date
      2009-03-29
    • Related Report
      2008 Annual Research Report
  • [Presentation] Artificial Peptide Tag Recognition by Dinuclear Zinc Complex and Luminescence Switchins with Lanthanide Complex2008

    • Author(s)
      T. Hirayama, M. Taki, Y. Yamamoto
    • Organizer
      The 4th Asian Biological Inorganic Chemistry Conference
    • Place of Presentation
      Jeju, Korea
    • Year and Date
      2008-11-12
    • Related Report
      2008 Annual Research Report
  • [Presentation] 蛍光性希土類錯体を用いた新しいペプチドタグープローブシステムの開発2008

    • Author(s)
      平山祐・多喜正泰・山本行男
    • Organizer
      第3回バイオ関連化学合同シンポジウム
    • Place of Presentation
      東京工業大学
    • Year and Date
      2008-09-20
    • Related Report
      2008 Annual Research Report 2008 Final Research Report
  • [Presentation] 非天然ペプチドタグを認識する二核亜鉛含有希土類錯体の開発2008

    • Author(s)
      平山祐, 多喜正泰, 山本行男
    • Organizer
      第58回錯体化学討論会
    • Place of Presentation
      金沢大学
    • Year and Date
      2008-09-19
    • Related Report
      2008 Annual Research Report
  • [Presentation] 希土類錯体を用いたペプチドタグ蛍光認識システムの開発2008

    • Author(s)
      平山祐・多喜正泰・山本行男
    • Organizer
      第21回配位化合物の光化学討論会
    • Place of Presentation
      北里大学
    • Year and Date
      2008-08-04
    • Related Report
      2008 Annual Research Report 2008 Final Research Report
  • [Presentation] 外部配位子との配位挙動に及ぼす環状ニッケル錯体の置換基効果2008

    • Author(s)
      若松俊彦・多喜正泰・山本行男
    • Organizer
      日本化学会第88春季年会
    • Place of Presentation
      立教大学
    • Year and Date
      2008-03-30
    • Related Report
      2007 Annual Research Report
  • [Presentation] タンパク質の可逆的蛍光ラベル化を指向した二官能基性ローダミン分子の開発2008

    • Author(s)
      春木秀仁・平山祐・多喜正泰・山本行男
    • Organizer
      日本化学会第88春季年会
    • Place of Presentation
      立教大学
    • Year and Date
      2008-03-28
    • Related Report
      2007 Annual Research Report
  • [Presentation] エチレングリコール担持金クラスターによる水銀イオンの特異的検出とその反応機構2008

    • Author(s)
      平山祐・多喜正泰・山本行男
    • Organizer
      日本化学会第88春季年会
    • Place of Presentation
      立教大学
    • Year and Date
      2008-03-28
    • Related Report
      2007 Annual Research Report
  • [Presentation] チオエーテル配位子を有するロサミン型蛍光センサーの構造と機能2008

    • Author(s)
      伊吉祥平・多喜正泰・山本行男
    • Organizer
      日本化学会第88春季年会
    • Place of Presentation
      立教大学
    • Year and Date
      2008-03-27
    • Related Report
      2007 Annual Research Report
  • [Presentation] Artificial Peptide Tag Recognition by Dinuclear Zinc Complex and Luminescence Switching with Lanthanide Complex2008

    • Author(s)
      Tasuku Hirayama, Masayasu Taki, Yukio Yamamoto
    • Organizer
      The 4th Asian Biological Inorganic Chemistry Conference
    • Place of Presentation
      Jeju, Korea
    • Related Report
      2008 Final Research Report
  • [Presentation] Development of New Lanthanide Complexes Binding to Peptide Tag2007

    • Author(s)
      Tasuku Hirayama, Masayasu Taki, Yukio Yamamoto
    • Organizer
      Biomimetics Conference
    • Place of Presentation
      Doshisha, Kyoto, Japan
    • Year and Date
      2007-12-20
    • Related Report
      2008 Final Research Report
  • [Presentation] Membrane-permeable Fluorescent Chemosensor for Ratiometric Imaging of Cadmium in2007

    • Author(s)
      Masayasu Taki and Yukio Yamamoto
    • Organizer
      Biomimetics Conference
    • Place of Presentation
      Doshisha University
    • Year and Date
      2007-12-08
    • Related Report
      2007 Annual Research Report
  • [Presentation] Development of New Lanthanide Complexes Binding to Peptide Tag2007

    • Author(s)
      Tasuku Hirayama, Masayasu Taki, Yukio Yamamoto
    • Organizer
      Biomimetics Conference
    • Place of Presentation
      Doshisha University
    • Year and Date
      2007-12-08
    • Related Report
      2007 Annual Research Report
  • [Presentation] 新しいペプチドタグ結合性希土類錯体の開発2007

    • Author(s)
      平山祐・多喜正泰・山本行男
    • Organizer
      第22回生体機能関連化学シンポジウム
    • Place of Presentation
      東北大学
    • Year and Date
      2007-09-28
    • Related Report
      2008 Final Research Report 2007 Annual Research Report
  • [Presentation] アミノクマリンを骨格とした新規レシオ型カドミウム蛍光プローブの開発2007

    • Author(s)
      多喜正泰・出崎美佳・山本行男
    • Organizer
      第22回生体機能関連化学シンポジウム
    • Place of Presentation
      東北大学
    • Year and Date
      2007-09-28
    • Related Report
      2007 Annual Research Report
  • [Presentation] エチレングリコール担持金クラスターを用いた水銀の特異的検出とその反応メカニズム2007

    • Author(s)
      平山祐・多喜正泰・山本行男
    • Organizer
      第57回錯体化学討論会
    • Place of Presentation
      名古屋工業大学
    • Year and Date
      2007-09-26
    • Related Report
      2007 Annual Research Report
  • [Presentation] チオエーテル配位子を有するロサミン型蛍光プローブの開発と金属イオンに対する蛍光挙動2007

    • Author(s)
      伊吉祥平・多喜正泰・山本行男
    • Organizer
      第57回錯体化学討論会
    • Place of Presentation
      名古屋工業大学
    • Year and Date
      2007-09-26
    • Related Report
      2007 Annual Research Report
  • [Presentation] タンパク質蛍光ラベル化を目指したペプチド結合性ニッケル錯体の合成と性質2007

    • Author(s)
      若松俊彦・平山祐・多喜正泰・山本行男
    • Organizer
      第57回錯体化学討論会
    • Place of Presentation
      名古屋工業大学
    • Year and Date
      2007-09-26
    • Related Report
      2007 Annual Research Report
  • [Presentation] Highly Selective Ratiometric Fluorescent Probe with Picomolar sensitivity for Cadmium2007

    • Author(s)
      Masayasu Taki and Yukio Yamamoto
    • Organizer
      13th International Conference on Biological Inorganic Chemistry
    • Place of Presentation
      University of Vienna
    • Year and Date
      2007-07-16
    • Related Report
      2007 Annual Research Report
  • [Presentation] Mechanism of colorimetric sensing of Hg2+ with triethyleneglycol-supported gold nano-cluster2007

    • Author(s)
      Tasuku Hirayama, Masayasu Taki, Yukio Yamamoto
    • Organizer
      13th International Conference on Biological Inorganic Chemistry
    • Place of Presentation
      University of Vienna
    • Year and Date
      2007-07-16
    • Related Report
      2007 Annual Research Report

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Published: 2007-04-01   Modified: 2016-04-21  

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