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Probing conformational changes in protein reaction sites via Raman optical activity

Research Project

Project/Area Number 21K04997
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Review Section Basic Section 32010:Fundamental physical chemistry-related
Research InstitutionSaga University

Principal Investigator

Fujisawa Tomotsumi  佐賀大学, 理工学部, 准教授 (60633493)

Project Period (FY) 2021-04-01 – 2024-03-31
Project Status Completed (Fiscal Year 2023)
Budget Amount *help
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2023: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2022: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2021: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Keywordsラマン光学活性 / 光受容タンパク質 / 低温分光 / 低温ラマン光学活性分光 / ラマン光学活性分光法 / 低温ラマン分光法 / 低温分光法 / 光受容体
Outline of Research at the Start

生体内で働くタンパク質はアミノ酸が立体的に連なった高分子であり、タンパク質の中には色素を内包したものが身近に存在する。例えば動物の目の中の光受容体は、タンパク質の中の色素が光を吸収して、その光化学反応によって最終的に光感知信号が導かれる。動物の視覚に代表されるこのような化学反応は、立体的なタンパク質の中で起こる、立体的な色素分子の化学変化である。しかし、今日でもタンパク質の中で反応する分子の3次元的な姿(構造)を正確に捉える技術は未熟である。本研究では、分子の3次元的構造に鋭敏なラマン光学活性分光法を用いて、生物の光受容体の中で反応する分子の立体構造を明らかにする。

Outline of Final Research Achievements

It is proteins that perform diverse chemical reactions in living organisms. In proteins, the visualization of three-dimensional structures of reacting molecules is one of the most important challenge. This research aimed to build up a new method using Raman optical activity (ROA) spectroscopy, which is very sensitive to three-dimensional (3D) structure of molecules, to track the biochemical reactions.
We chose the photoreceptors of micro-organisms, such as microbial rhodopsins and photoactive yellow proteins, for the target of our study and planned to measure ROA of their photointermediates trapped at low temperature. As the first step for this ROA measurement, we measured the low-temperature Raman spectra of the photointermediates of various photoreceptors and analyzed their structural changes in the photoreactions. Then, we extended our this Raman spectrometer to the low-temperature ROA spectrometer to obtain the three-dimensional structure information of photointermediates.

Academic Significance and Societal Importance of the Research Achievements

本研究で実施した光受容タンパク質の低温ラマン測定は良質な分子のスペクトルを得られる方法でありながら、世界的にみても同様の手法を用いる研究グループがほとんど無い。そのため、独自性ある構造情報に基づいた研究成果を発信することができる。さらに、これを発展させた低温ラマン光学活性(ROA)分光法は、今日広く利用されるどの分光手法でも測定ができない、溶液中のタンパク質の反応サイトの立体構造変化を捉えられる方法となる。本研究はタンパク質研究において"今まで見えなかった構造を観測する"展開を開拓する取り組みである

Report

(4 results)
  • 2023 Annual Research Report   Final Research Report ( PDF )
  • 2022 Research-status Report
  • 2021 Research-status Report
  • Research Products

    (42 results)

All 2024 2023 2022 2021 Other

All Int'l Joint Research (5 results) Journal Article (12 results) (of which Int'l Joint Research: 3 results,  Peer Reviewed: 12 results,  Open Access: 4 results) Presentation (21 results) (of which Int'l Joint Research: 5 results,  Invited: 4 results) Book (1 results) Remarks (3 results)

  • [Int'l Joint Research] オクラホマ州立大学(米国)

    • Related Report
      2023 Annual Research Report
  • [Int'l Joint Research] チェコ科学アカデミー(チェコ)

    • Related Report
      2023 Annual Research Report
  • [Int'l Joint Research] オクラハマ州立大学(米国)

    • Related Report
      2022 Research-status Report
  • [Int'l Joint Research] チェコ科学アカデミー(チェコ)

    • Related Report
      2022 Research-status Report
  • [Int'l Joint Research] オクラハマ州立大学(米国)

    • Related Report
      2021 Research-status Report
  • [Journal Article] Near-Infrared Excited Raman Optical Activity as a Tool to Uncover Active Sites of Photoreceptor Proteins2024

    • Author(s)
      Fujisawa Tomotsumi、Unno Masashi
    • Journal Title

      The Journal of Physical Chemistry B

      Volume: 128 Issue: 10 Pages: 2228-2235

    • DOI

      10.1021/acs.jpcb.4c00094

    • Related Report
      2023 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Configurational Changes of Retinal Schiff Base during Membrane Na<sup>+</sup> Transport by a Sodium Pumping Rhodopsin2024

    • Author(s)
      Fujisawa Tomotsumi、Kinoue Kouta、Seike Ryouhei、Kikukawa Takashi、Unno Masashi
    • Journal Title

      The Journal of Physical Chemistry Letters

      Volume: 15 Issue: 7 Pages: 1993-1998

    • DOI

      10.1021/acs.jpclett.3c03435

    • Related Report
      2023 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Molecular Vibrations in Chiral Europium Complexes Revealed by Near‐Infrared Raman Optical Activity2023

    • Author(s)
      Wu Tao、Bour Petr、Fujisawa Tomotsumi、Unno Masashi
    • Journal Title

      Advanced Science

      Volume: 11 Issue: 1 Pages: 2305521-2305521

    • DOI

      10.1002/advs.202305521

    • Related Report
      2023 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Spectroscopic Validation of Crystallographic Structures of a Protein Active Site by Chiroptical Spectroscopy2023

    • Author(s)
      Fujisawa Tomotsumi、Shingae Takahito、Ren Jie、Haraguchi Shojiro、Hanamoto Takeshi、Hoff Wouter D.、Unno Masashi
    • Journal Title

      The Journal of Physical Chemistry Letters

      Volume: 14 Issue: 41 Pages: 9304-9309

    • DOI

      10.1021/acs.jpclett.3c01954

    • Related Report
      2023 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Raman Optical Activity of Retinal Chromophore in Sensory Rhodopsin II2023

    • Author(s)
      Nishikawa Kouhei、Kuroiwa Ryosuke、Tamogami Jun、Unno Masashi、Fujisawa Tomotsumi
    • Journal Title

      The Journal of Physical Chemistry B

      Volume: 127 Issue: 33 Pages: 7244-7250

    • DOI

      10.1021/acs.jpcb.3c02391

    • Related Report
      2023 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Reisomerization of retinal represents a molecular switch mediating Na+ uptake and release by a bacterial sodium-pumping rhodopsin2022

    • Author(s)
      Fujisawa, T., Kinoue, K. Seike, R., Kikukawa, T., Unno, M.
    • Journal Title

      Journal of biological chemistry

      Volume: 298 Issue: 9 Pages: 102366-102366

    • DOI

      10.1016/j.jbc.2022.102366

    • Related Report
      2022 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Conformation of Retinal Schiff Base in Proteorhodopsin Based on Raman Optical Activity2022

    • Author(s)
      藤澤知績、海野雅司
    • Journal Title

      Seibutsu Butsuri

      Volume: 62 Issue: 6 Pages: 331-333

    • DOI

      10.2142/biophys.62.331

    • ISSN
      0582-4052, 1347-4219
    • Related Report
      2022 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] 近赤外ラマン分光法の色素タンパク質への応用2022

    • Author(s)
      藤澤知績、海野雅司
    • Journal Title

      分光研究

      Volume: 71 Pages: 105-107

    • Related Report
      2022 Research-status Report
    • Peer Reviewed
  • [Journal Article] Raman Spectroscopy of an Atypical C15-E,syn Bilin Chromophore in Cyanobacteriochrome RcaE2022

    • Author(s)
      Yuji Okuda, Risako Miyoshi, Takanari Kamo, Tomotsumi Fujisawa, Takayuki Nagae, Masaki Mishima, Toshihiko Eki, Yuu Hirose, Masashi Unno
    • Journal Title

      J. Phys. Chem. B

      Volume: 126 Issue: 4 Pages: 813-821

    • DOI

      10.1021/acs.jpcb.1c09652

    • Related Report
      2021 Research-status Report
    • Peer Reviewed
  • [Journal Article] Femtosecond time-resolved absorption study of signaling state of a BLUF protein PixD from the cyanobacterium synechocystis: hydrogen bond rearrangement completes during forward proton-coupled electron transfer2021

    • Author(s)
      T. Fujisawa, S. Masuda, S. Takeuchi, T. Tahara
    • Journal Title

      Journal of Physical Chemistry B

      Volume: 125 Issue: 44 Pages: 12154-12165

    • DOI

      10.1021/acs.jpcb.1c05957

    • Related Report
      2021 Research-status Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Conformational analysis of a retinal Schiff base chromophore in proteorhodopsin by Raman optical activity2021

    • Author(s)
      Fujisawa, T., Nishikawa, K., Tamogami, J., Unno, M.
    • Journal Title

      J. Phys. Chem. Lett.

      Volume: 12 Issue: 39 Pages: 9564-9568

    • DOI

      10.1021/acs.jpclett.1c02552

    • Related Report
      2021 Research-status Report
    • Peer Reviewed
  • [Journal Article] Structural basis of the protochromic green/red photocycle of the chromatic acclimation sensor RcaE2021

    • Author(s)
      Takayuki Nagae, Masashi Unno, Taiki Koizumi, Yohei Miyanoiri, Tomotsumi Fujisawa, Kento Masui, Takanari Kamo, Kei Wada, Toshihiko Eki, Yutaka Ito, Yuu Hirose, Masaki Mishima
    • Journal Title

      Proceedings of the National Academy of Sciences

      Volume: 118(20) Issue: 20

    • DOI

      10.1073/pnas.2024583118

    • Related Report
      2021 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] Raman optical activity spectroscopy of biological photoreceptors2024

    • Author(s)
      Tomotsumi Fujisawa
    • Organizer
      24th East Asian Workshop on Chemical Dynamics (EAWCD 2024)
    • Related Report
      2023 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Structure and Heterogeneity of Retinal Chromophore in Chloride Pump Rhodopsins Revealed by Raman Optical Activity2023

    • Author(s)
      Masashi Unno, Masaiku Ohya, Takashi Kikukawa, Junpei Matsuo, Takashi Tsukamoto, Ryota Nagaura, Tomotsumi Fujisawa
    • Organizer
      第61回日本生物物理学会年会
    • Related Report
      2023 Annual Research Report
  • [Presentation] Configurational change of retinal Schiff base chromophore in a sodium pump rhodopsin2023

    • Author(s)
      Tomotsumi Fujisawa, Kouta Kinoue, Ryouhei Seike, Takashi Kikukawa, Masashi Unno
    • Organizer
      第61回日本生物物理学会年会
    • Related Report
      2023 Annual Research Report
  • [Presentation] Structural analysis of the blue-absorbing form of blue/orange cyanobacteriochrome by Raman spectroscopy2023

    • Author(s)
      Ryoka Seto, Masako Hamada, Yuu Hirose, Tomotsumi Fujisawa, Masashi Unno
    • Organizer
      第61回日本生物物理学会年会
    • Related Report
      2023 Annual Research Report
  • [Presentation] Role of Lys261 in Cyanobacteriochrome RcaE Studied by Mutagenesis and Raman Spectroscopy2023

    • Author(s)
      Taisei Koga, Masako Hamada, Yuu Hirose, Tomotsumi Fujisawa, Masashi Unno
    • Organizer
      第61回日本生物物理学会年会
    • Related Report
      2023 Annual Research Report
  • [Presentation] Low-temperature spectroscopy of photoactive yellow protein from Rhodospirillum centenum2023

    • Author(s)
      Kirari Ogata, Tomotsumi Fujisawa, Wouter D Hoff, Masashi Unno
    • Organizer
      第61回日本生物物理学会年会
    • Related Report
      2023 Annual Research Report
  • [Presentation] ラマン光学活性分光法を用いた センサリーロドプシンⅡの発色団コンフォメーションの研究2023

    • Author(s)
      藤澤知績,西川航平,黒岩亮介,田母神淳,海野雅司
    • Organizer
      第45回溶液化学シンポジウム
    • Related Report
      2023 Annual Research Report
  • [Presentation] 低温ラマン分光法による Photoactive Yellow Proteinの L中間体の構造解析2023

    • Author(s)
      川崎翔太,藤澤知績, 海野雅司, Wouter D. Hoff
    • Organizer
      第45回溶液化学シンポジウム
    • Related Report
      2023 Annual Research Report
  • [Presentation] Photoconversion mechanism of a green/red cyanobacteriochrome based on its molecular structure2023

    • Author(s)
      Tomotsumi Fujisawa
    • Organizer
      15th International Conference on Tetrapyrrole Photoreceptors in Photosynthetic Organisms
    • Related Report
      2023 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Cryogenic Raman study of photoconversion from green to red absorbing state of the cyanobacteriochrome RcaE2023

    • Author(s)
      Yasuhiro Jyojima, Masako Hamada, Yuu Hirose, Masashi Unno, Tomotsumi Fujisawa
    • Organizer
      15th International Conference on Tetrapyrrole Photoreceptors in Photosynthetic Organisms
    • Related Report
      2023 Annual Research Report
    • Int'l Joint Research
  • [Presentation] クライオ時間分解ラマン分光法によるナトリウムポンプロドプシンの研究: O中間体における発色団構造変化2023

    • Author(s)
      藤澤知績,木ノ上 晃汰,清家陵平,菊川 峰志,海野雅司
    • Organizer
      第17回分子科学討論会
    • Related Report
      2023 Annual Research Report
  • [Presentation] Cryogenic Raman study of primary photointermediate of photoactive yellow protein2023

    • Author(s)
      Tomotsumi Fujisawa, Daisuke Shiga, Akihiro Baba, Wouter D. Hoff, Masashi Unno
    • Organizer
      8th Asian Spectroscopy Conference
    • Related Report
      2023 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Raman optical activity spectroscopy for measuring chromophore conformations in biological photoreceptors2023

    • Author(s)
      Tomotsumi Fujisawa
    • Organizer
      3rd Q-PIT photofunction integration symposium
    • Related Report
      2023 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Time-resolved cryo-Raman study of Na+ uptake and release by a sodium pumping rhodopsin2022

    • Author(s)
      Tomotsumi Fujisawa, Kouta Kinoue, Ryouhei Seike, Takashi Kikukawa, Masashi Unno
    • Organizer
      第60回生物物理学会年会
    • Related Report
      2022 Research-status Report
  • [Presentation] 低温ラマン分光法によるphotoactive yellow proteinのL中間体の構造解析2022

    • Author(s)
      Shota Kawasaki, Tomotsumi Fujisawa, Wouter D. Hoff, Masashi Unno
    • Organizer
      第60回生物物理学会年会
    • Related Report
      2022 Research-status Report
  • [Presentation] シアノバクテリオクロムRcaEにおけるユニークなC15-E, syn型ビリン発色団のラマン分光法による研究2022

    • Author(s)
      Yuji Okuda, Risako Miyoshi, Takanari Kamo, Tomotsumi Fujisawa, Takayuki Nagae, Masaki Mishima, Toshihiko Eki, Yuu Hirose, Masashi Unno
    • Organizer
      第60回生物物理学会年会
    • Related Report
      2022 Research-status Report
  • [Presentation] 低温ラマン分光法による光駆動Cl-ポンプNMR3の発色団構造変化の研究2022

    • Author(s)
      Natsumi Ejima, Tomotsumi Fujisawa, Takashi Kikukawa, Masashi Unno
    • Organizer
      第60回生物物理学会年会
    • Related Report
      2022 Research-status Report
  • [Presentation] 光駆動クロライドイオンポンプロドプシンのラマン分光法を用いた構造解析2022

    • Author(s)
      江嶌那月,藤澤知績,菊川峰志,海野雅司
    • Organizer
      溶液化学研究会若手の会 第2回冬季発表会
    • Related Report
      2022 Research-status Report
  • [Presentation] 低温ラマン分光法による Photoactive Yellow Protein の初期中間体の構造決定2021

    • Author(s)
      志賀 大将 , 藤澤 知績2, Hoff Wouter D. , 海野 雅司
    • Organizer
      第59回日本生物物理学会年会
    • Related Report
      2021 Research-status Report
  • [Presentation] ラマン光学活性による微生物型ロドプシン単量体と多量体におけるレチナールシッフ塩基の構 造解析2021

    • Author(s)
      西川 航平 , 藤澤 知績 2, 海野 雅司 , 須藤 雄気 , 小島 慧一 , 仲間 政樹
    • Organizer
      第59回日本生物物理学会年会
    • Related Report
      2021 Research-status Report
  • [Presentation] ラマン光学活性によるセンサリーロドプシンⅡのレチナールシッフ塩基の構造解析2021

    • Author(s)
      西川航平,田母神淳,藤澤知績,海野雅司
    • Organizer
      溶液化学研究会若手の会第1回冬季発表会
    • Related Report
      2021 Research-status Report
  • [Book] Advanced spectral analysis of complex molecular system; In Molecular and Laser Spectroscopy: Advances and Applications: Volume 32022

    • Author(s)
      Fujisawa, T., Unno, M.
    • Total Pages
      30
    • Publisher
      Elsevier
    • ISBN
      9780128498828
    • Related Report
      2022 Research-status Report
  • [Remarks] 佐賀大学 海野-藤澤研究室

    • URL

      http://biophysics.chem.saga-u.ac.jp/unno-lab/homu.html

    • Related Report
      2023 Annual Research Report
  • [Remarks] 藤澤知績、海野‐藤澤研究室

    • URL

      http://biophysics.chem.saga-u.ac.jp/Fujisawa_webpage/Tfujisawa.html

    • Related Report
      2022 Research-status Report
  • [Remarks] 佐賀大学 海野‐藤澤研究室

    • URL

      http://biophysics.chem.saga-u.ac.jp/unno-lab/homu.html

    • Related Report
      2021 Research-status Report

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Published: 2021-04-28   Modified: 2025-01-30  

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