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Search for assembly intermediates of photosynthetic proteins by high-throughput single-molecule spectroscopy

Research Project

Project/Area Number 19H03187
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Review Section Basic Section 43040:Biophysics-related
Research InstitutionTohoku University

Principal Investigator

Shibata Yutaka  東北大学, 理学研究科, 准教授 (20300832)

Co-Investigator(Kenkyū-buntansha) 高橋 裕一郎  岡山大学, 異分野基礎科学研究所, 教授 (50183447)
鞆 達也  東京理科大学, 教養教育研究院神楽坂キャンパス教養部, 教授 (60300886)
Project Period (FY) 2019-04-01 – 2022-03-31
Project Status Completed (Fiscal Year 2021)
Budget Amount *help
¥15,730,000 (Direct Cost: ¥12,100,000、Indirect Cost: ¥3,630,000)
Fiscal Year 2021: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
Fiscal Year 2020: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
Fiscal Year 2019: ¥10,140,000 (Direct Cost: ¥7,800,000、Indirect Cost: ¥2,340,000)
Keywords低温顕微鏡 / タンパク質Assembly / 励起スペクトル / 単一分子分光 / 時間分解蛍光測定 / エネルギー移動 / 過渡的中間体 / 光化学系I / 過渡的中華体
Outline of Research at the Start

植物の光合成を駆動するのは、数10個の色素分子を結合したタンパク質複合体である。その複雑な構造が生体内で構築される機構はよく分かっていない。これを明らかにするため、本研究では構造が出来上がる途上の中間体の同定を目指す。光をあてずに発芽させた植物に光照射すると、活発な光合成タンパク質合成が開始される。このような植物に含まれる光合成タンパク質を丸ごと全部溶液中に溶かしで希釈し、単一分子分光という方法で一つ一つのタンパク質の蛍光スペクトルを取得する。最近私が開発した顕微鏡では、多くの分子のスペクトルを効率よく取得できるようになっており、この技術を利用して微量に存在する構造構築中間体を見つける。

Outline of Final Research Achievements

In order to reveal the mechanism based on which photosynthetic proteins are automatically assembled within biological systems, I aimed to detect the intermediate states of their assembly processes. Since such intermediates are generated only transiently and therefore accumulate only a tiny amount, detection of these components are quite difficult. To overcome the difficulty, I extended the single molecule detection technique to perform an exhaustive survey of all the components within a sample. The present development have established a method to characterize components with a tiny amount within a sample.

Academic Significance and Societal Importance of the Research Achievements

光合成を駆動するタンパク質の構造は明らかとなり、反応機構解明も進んだ。一方で、複雑な構造が生体内で自発的に構築される機構はほとんど分かっていない。特に100個近い色素分子が、大きなタンパク質分子の所定の位置に正しく結合する機構は全くの謎である。光合成反応を駆動するタンパク質の構築機構が分かれば、人工的に光合成反応を起こす分子機械の合成にもヒントとなると期待される。

Report

(4 results)
  • 2021 Annual Research Report   Final Research Report ( PDF )
  • 2020 Annual Research Report
  • 2019 Annual Research Report
  • Research Products

    (22 results)

All 2021 2020 2019 Other

All Journal Article (6 results) (of which Peer Reviewed: 6 results,  Open Access: 2 results) Presentation (15 results) (of which Int'l Joint Research: 2 results) Remarks (1 results)

  • [Journal Article] High-speed excitation-spectral microscopy uncovers in-situ rearrangement of light-harvesting apparatus in Chlamydomonas during state transitions at submicron precision2021

    • Author(s)
      Zhang XianJun、Fujita Yuki、Tokutsu Ryutaro、Minagawa Jun、Ye Shen、Shibata Yutaka
    • Journal Title

      Plant and Cell Physiology

      Volume: 未定 Issue: 5 Pages: 872-882

    • DOI

      10.1093/pcp/pcab047

    • Related Report
      2021 Annual Research Report 2020 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Fluctuating Energy-Transfer Pathway of Photosynthetic Antenna Systems Observed by Single-Molecule Fluorescence Spectroscopy2021

    • Author(s)
      SHIBATA Yutaka
    • Journal Title

      Seibutsu Butsuri

      Volume: 61 Issue: 1 Pages: 023-026

    • DOI

      10.2142/biophys.61.023

    • NAID

      130007976661

    • ISSN
      0582-4052, 1347-4219
    • Related Report
      2021 Annual Research Report 2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Development of a multicolor line-focus microscope for rapid acquisitions of excitation spectra2020

    • Author(s)
      Sakar Jana, and Yutaka Shibata
    • Journal Title

      Biophysical Journal

      Volume: 118 Issue: 1 Pages: 36-43

    • DOI

      10.1016/j.bpj.2019.11.023

    • Related Report
      2020 Annual Research Report 2019 Annual Research Report
    • Peer Reviewed
  • [Journal Article] A gold nanoparticle conjugate with photosystem I and photosystem II for development of a biohybrid water-splitting photocatalyst2020

    • Author(s)
      Kawahara Kousuke、Inoue-Kahino Natsuko、Namie Keisuke、Kato Yuki、Tomo Tatsuya、Shibata Yutaka、Kashino Yasuhiro、Noguchi Takumi
    • Journal Title

      Biomedical Spectroscopy and Imaging

      Volume: 9 Issue: 1-2 Pages: 73-81

    • DOI

      10.3233/bsi-200200

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Identification of Assembly Precursors to Photosystems Emitting Fluorescence at 683 nm and 687 nm by Cryogenic Fluorescence Microspectroscopy2019

    • Author(s)
      Tomofumi Chiba, and Yutaka Shibata
    • Journal Title

      Biochim. Biophys. Acta

      Volume: 1860 Issue: 12 Pages: 148090-148090

    • DOI

      10.1016/j.bbabio.2019.148090

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Redox-state dependent blinking of single photosystem I trimers at around liquid-nitrogen temperature2019

    • Author(s)
      Jana, S., Du, T., Nagao, R., Noguchi, T., Shibata, Y.
    • Journal Title

      Biochim. Biophys. Acta, Bioenerg.

      Volume: 1860 Issue: 1 Pages: 30-40

    • DOI

      10.1016/j.bbabio.2018.11.002

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed
  • [Presentation] Excitation-spectral microscopy uncovers in-situ rearrangement of LHCII in Chlamydomonas during state transitions2021

    • Author(s)
      張 先駿, 藤田 祐輝, 得津 隆太郎, 皆川 純, 叶 深, 柴田 穣
    • Organizer
      第11回日本光合成学会年会
    • Related Report
      2021 Annual Research Report
  • [Presentation] Excitation-spectral microscopy uncovers in-situ rearrangement of LHCII in Chlamydomonas during state transition2021

    • Author(s)
      張 先駿, 藤田 祐輝, 得津 隆太郎, 皆川 純, 叶 深, 柴田 穣
    • Organizer
      第28回光合成セミナー2021: 反応中心と色素系の多様性
    • Related Report
      2021 Annual Research Report
  • [Presentation] 光化学系Iassembly中間体の時間分解蛍光および単一分子分光解析2021

    • Author(s)
      金田直也, Sreedhar Nellaepalli, 高橋裕一郎, 柴田穣
    • Organizer
      第28回光合成セミナー2021: 反応中心と色素系の多様性
    • Related Report
      2021 Annual Research Report
  • [Presentation] 二次元蛍光寿命相関分光による光化学系Iタンパク質の単一分子電子移動速度の測定2021

    • Author(s)
      谷口凜,篠田稔行,鞆達也,叶深,近藤徹,柴田穣
    • Organizer
      第15回分子科学討論会
    • Related Report
      2021 Annual Research Report
  • [Presentation] 光化学系I assembly中間体の単粒子分光と時間分解蛍光解析2021

    • Author(s)
      金田直也, Sreedhar Nellaepalli, 高橋裕一郎, 叶深, 柴田穣
    • Organizer
      2021年度 化学系学協会東北大会
    • Related Report
      2021 Annual Research Report
  • [Presentation] ストリークカメラを検出器とした細胞内局所での時間分解顕微蛍光分光2021

    • Author(s)
      藤田 祐輝,張 先駿, 金田 直也,柴田 穣
    • Organizer
      第59回日本生物物理学会年会
    • Related Report
      2021 Annual Research Report
  • [Presentation] 励起スペクトル顕微鏡と超解像イメージングで明らかになったステート遷移におけるチラコイ ド膜の不規則な構造変化2021

    • Author(s)
      張 先駿, 藤田 祐輝, 金田 直也, 得津 隆太郎, 皆川 純, 叶 深, 柴田 穣
    • Organizer
      第59回日本生物物理学会年会
    • Related Report
      2021 Annual Research Report
  • [Presentation] Uphill energy transfer mechanism for photosynthesis in the Antarctic alga2021

    • Author(s)
      Makiko Kosugi, Masato Kawasaki, Yutaka Shibata, Kojiro Hara, Shinichi Takaichi, Toshio Moriya, Naruhiko Adachi, Yasuhiro Kamei, Yasuhiro Kashino, Sakae Kudoh, Hiroyuki Koike, Toshiya Senda
    • Organizer
      The 12th Symposium on Polar Science
    • Related Report
      2021 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Connecting the spectral properties to the structure of photosystem I containing Chlorophyll-f2020

    • Author(s)
      Rin Taniguchi, Yutaka Shibata, Toshiyuki Shinoda, Tatsuya Tomo, Shen Ye
    • Organizer
      第58回日本生物物理学会年会
    • Related Report
      2020 Annual Research Report
  • [Presentation] In situ visualization of reversible state transition in live Chlamydomonas cells by noninvasive excitation spectral microscopy2020

    • Author(s)
      Xianjun Zhang, Yuki Fujita, Ryutaro Tokutsu, Jun Minagawa, Shen Ye, Yutaka Shibata
    • Organizer
      第58回日本生物物理学会年会
    • Related Report
      2020 Annual Research Report
  • [Presentation] Simultaneous Mapping of Fluorescence Spectra and Lifetimes of Chlorophylls Revealed Accumulation of Quenched LHCII2020

    • Author(s)
      Yuki Fujita, Xianjun Zhang, Touru Kondou, Yutaka Shibata
    • Organizer
      第58回日本生物物理学会年会
    • Related Report
      2020 Annual Research Report
  • [Presentation] 単一分子分光によるクロロフィル-f を含む光化学系I の蛍光不均一性2020

    • Author(s)
      小林 誉宗,谷口 凛,柴田 穣,篠田 稔行,鞆 達也,叶 深
    • Organizer
      第61回日本植物生理学会年会
    • Related Report
      2019 Annual Research Report
  • [Presentation] 極低温光学顕微分光法による緑藻クラミドモナスのステート遷移の葉緑体内不均一性の観測2020

    • Author(s)
      藤田祐輝,張先駿,柴田穣
    • Organizer
      第61回日本植物生理学会年会
    • Related Report
      2019 Annual Research Report
  • [Presentation] Single Molecule Spectroscopy On Photosystem I Revealed The Dynamical Nature Of The Light Harvesting2019

    • Author(s)
      Yutaka Shibata, Sankar Jana, Ting Du, Ryo Nagao, and Takumi Noguchi
    • Organizer
      International Conference on Artificial Photosynthesis-2019
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Evaluation of the movement of photosynthetic antenna complexes in algal cells by excitation spectral measurements on microscope2019

    • Author(s)
      Xianjun Zhang, Sankar Jana, Shen Ye, Yutaka Shibata
    • Organizer
      International Conference on Artificial Photosynthesis-2019
    • Related Report
      2019 Annual Research Report
  • [Remarks] 光で単一分子の挙動を測る

    • URL

      http://sub.web.tohoku.ac.jp/orgphys/content/files/research_Shibata.pdf

    • Related Report
      2021 Annual Research Report

URL: 

Published: 2019-04-18   Modified: 2023-01-30  

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