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2010 Fiscal Year Annual Research Report

変異体構造解析による光化学系II複合体の構造制御機構の解明

Research Project

Project/Area Number 20570038
Research InstitutionOkayama University

Principal Investigator

沈 建仁  岡山大学, 大学院・自然科学研究科, 教授 (60261161)

Keywords光合成 / 光化学系II / 膜タンパク質 / 結晶化 / 結晶構造解析 / タンパク質複合体 / 酸素発生 / 変異体
Research Abstract

本研究の目的は、光合成光化学系II複合体(以下PSIIとする)における各低分子量サブユニットの役割を、それぞれのサブユニット欠失変異体を用いて、結晶化・X線結晶構造解析と機能解析を組み合わせて解析し、PSII複合体の構造形成・制御機構を明らかにことである。これまで好熱性シアノバクテリアThermosynechococcus vulcanusを用いて、PsbM,PsbI,PsbZ,Ycfl2の各サブユニット欠失変異株からPSIIを精製・結晶化し、その結晶構造解析に成功し、立体構造に基づき各サブユニットの機能に関する新しい知見を得た。しかし、これまでの結晶の分解能が十分高くなく、また、低分子量サブユニットの欠失によるPSIIの構造変化が小さいため、各々のサブユニットの欠失によるPSIIの構造変化を明らかにすることはできなかった。従って、結晶の分解能を向上させ、PSIIの構造をより高分解能で解析することが本研究の緊急な課題となった。これには、まず野生株由来PSIIの結晶を改善し、分解能を向上させる必要があった。そこで本年度では、野生株由来PSIIの結晶化条件、結晶の抗凍結剤条件などを改善し、結晶の分解能を従来の2.9-3.0 Aから1.9 Aに大幅に改善することができた。得られた結晶を用いて構造解析を行った結果、真の原子分解能でPSIIの構造を解析することができ、これまで未知であったMn4CaO5クラスターの構造や多くのアミノ酸、及び補欠因子の詳細な構造を解明することができた。これにより変異株におけるサブユニットの欠失による構造変化を解明する手段が確立された。同様の方法で変異株由来PSIIを結晶化し、高分解能の構造解析を行えば、野生株と変異株の構造的違いを明らかにすることができ、サブユニットの欠失によるPSIIの構造変化を明らかにすることができる。

  • Research Products

    (24 results)

All 2011 2010

All Journal Article (7 results) (of which Peer Reviewed: 7 results) Presentation (14 results) Book (3 results)

  • [Journal Article] Photosynthetic oxygen evolution in mesoporous silica material : Adsorption of photosystem II reaction center complex into 23-nm nanopores in SBA2011

    • Author(s)
      Tomoyasu Noji, Chihiro Kamidaki, Keisuke Kawakami, Jian-Ren Shen, Tsutomu Kajino, Yoshiaki Fukushima, Takeshi Sekitoh, Shigeru Itoh
    • Journal Title

      Langmuir

      Volume: 27 Pages: 705-713

    • Peer Reviewed
  • [Journal Article] Roles of PsbI and PsbM in photosystem II dimer formation and stability studied by deletion mutagenesis and X-ray crystallography2011

    • Author(s)
      Keisuke Kawakami, Yasufumi Umena, Masako Iwai, Yousuke Kawabata, Masahiko Ikeuchi.
    • Journal Title

      Biochim.Biophys.Acta

      Volume: 1807 Pages: 319-325

    • Peer Reviewed
  • [Journal Article] Proton-Coupled Electron-Transfer Processes in Photosystem II Probed by Highly Resolved g-Anisotropy of Redox-Active Tyrosine Y(Z)2011

    • Author(s)
      Matsuoka H., Shen J.-R.Kawamori A., Nishiyama K., Ohba Y., Yamauchi S.
    • Journal Title

      J.Am.Chem.Soc.

      Volume: 133 Pages: 4655-4660

    • Peer Reviewed
  • [Journal Article] Labile electronic and spin states of the CaMn405 cluster in the PSII system refined to the 1.9 A X-ray resolution. UB3LYP computational results2011

    • Author(s)
      Kanda K., Yamanaka S., Saito T., Umena Y., Kawakami K., Shen J.-R., Kamiya N., Okumura M., Nakamura H., Yamaguchi K.
    • Journal Title

      Chem.Phys.Lett.

      Volume: 506 Pages: 98-103

    • Peer Reviewed
  • [Journal Article] Structural and functional studies on Ycf12 (Psb30) and PsbZ deletion mutants from a thermophilic cyanobacterium2010

    • Author(s)
      Kenji Takasaka, Masako Iwai, Yasufumi Umena, Keisuke Kawakami, Yukari Ohmori.
    • Journal Title

      Biochim.Biophys.Acta

      Volume: 1797 Pages: 278-284

    • Peer Reviewed
  • [Journal Article] Topological analysis of the extrinsic PsbO, PsbP and PsbQ proteins in a green algal PSII complex by cross-linking with a water-soluble carbodiimide2010

    • Author(s)
      Nagao R, Suzuki T, Okumura A, Niikura A, Iwai M, Dohmae N, Tomo T, Shen J.-R, Ikeuchi M, Enami I.
    • Journal Title

      Plant Cell Physiol.

      Volume: 51 Pages: 718-727

    • Peer Reviewed
  • [Journal Article] Binding and functional properties of five extrinsic proteins in oxygen-evolving Photosystem II from a marine centric diatom. Chaetoceros gracilis2010

    • Author(s)
      Nagao R., Moriguchi A., Tomo T., Niikura A., Nakajima A., Suzuki T., Okumura A., Iwai M., Shen J.-R., Ikeuchi M., Enami I.
    • Journal Title

      J.Biol.Chem.

      Volume: 285 Pages: 29191-29199

    • Peer Reviewed
  • [Presentation] Crystal structure of oxygen evolving photosystem II at an atomic resolution2010

    • Author(s)
      Jian-Ren Shen, Yasufumi Umena, Keisuke Kawakami, Nobuo Kamiya
    • Organizer
      The 70th Okazaki International Conference on Molecular mechanism of photosynthetic energy conversion : the present
    • Place of Presentation
      Okazaki,Japan
    • Year and Date
      20101204-20101206
  • [Presentation] 光化学系II複合体結晶の分解能向上2010

    • Author(s)
      川上恵典, 梅名泰史, 田代隆慶, 神谷信夫, 沈建仁
    • Organizer
      日本結晶学会2010年度年会
    • Place of Presentation
      大阪
    • Year and Date
      20101203-20101205
  • [Presentation] 1.9Å分解能の光化学系II膜蛋白質複合体の結晶構造解析2010

    • Author(s)
      梅名泰史・川上恵典・沈建仁・神谷信夫
    • Organizer
      日本結晶学会2011年度年会
    • Place of Presentation
      大阪
    • Year and Date
      20101203-20101205
  • [Presentation] Crystal structure of oxygen evolving Photosystem II at atomic resolution2010

    • Author(s)
      Yasufumi Umena, Keisuke Kawakami, Jian-Ren Shen, Nobuo Kamiya
    • Organizer
      26th European Crystallographic Meeting
    • Place of Presentation
      Darmstadt, Germany
    • Year and Date
      20100829-20100902
  • [Presentation] Crystal structure of oxygen evolving Photosystem II at an atomic resolution2010

    • Author(s)
      Jian-Ren Shen, Yasufumi Umena, Keisuke Kawakami, Nobuo Kamiya
    • Organizer
      15th International Congress Of Photosynthesis (PS2010)
    • Place of Presentation
      Beijing, China
    • Year and Date
      20100822-20100828
  • [Presentation] Improvement of the quality of photosystem II crystals for its structural analysis at an atomic resolution2010

    • Author(s)
      Keisuke Kawakami, Yasufumi Umena, Takayoshi Tashiro, Nobuo Kamiya.
    • Organizer
      15th International Congress Of Photosynthesis (PS2010)
    • Place of Presentation
      Beijing, China
    • Year and Date
      20100822-20100828
  • [Presentation] Charactarization of photosystem 11 from extrinsic protein-deleted mutants of a thermophilic cyanobacterium2010

    • Author(s)
      Yukari Ohmori, Masako Iwai, Masahiko Ikeuchi, Jian-Ren Shen
    • Organizer
      15th International Congress Of Photosynthesis (PS2010)
    • Place of Presentation
      Beijing, China
    • Year and Date
      20100822-20100828
  • [Presentation] Purification and characterization of oxygen-evolving Photosystem II complex from a moderate thermophilic diatom, Chaetoceros neogracile2010

    • Author(s)
      Ayako Niikura, Ryo Nagao, Isao Enami, Jian-Ren Shen
    • Organizer
      15th International Congress Of Photosynthesis (PS2010)
    • Place of Presentation
      Beijing, China
    • Year and Date
      20100822-20100828
  • [Presentation] The Position and Orientation of Active Carotenoid in Photosystem II2010

    • Author(s)
      Asako Kawamori, Hiroyuki Mino, Jian-Ren Shen
    • Organizer
      15th International Congress Of Photosynthesis (PS2010)
    • Place of Presentation
      Beijing, China
    • Year and Date
      20100822-20100828
  • [Presentation] Photosynthetic oxygen evolution from PSII reaction centers in nanopores inside silica mesoporous material2010

    • Author(s)
      Tomoyasu Noji, Chihiro Kamidaki, Keisuke Kawakami, Jian-Ren Shen, Tsutomu Kajino, Yoshiaki Fukushima, Takeshi
    • Organizer
      15th International Congress Of Photosynthesis (PS2010)
    • Place of Presentation
      Beijing, China
    • Year and Date
      20100822-20100828
  • [Presentation] 紅藻由来光化学系II複合体の精製と晶の分解能の改善と構造解析の現状2010

    • Author(s)
      足立秀行、梅名泰史、田代隆慶、榎並勲、神谷信夫、沈建仁
    • Organizer
      第16回日本光生物学協会年会
    • Place of Presentation
      大阪
    • Year and Date
      20100810-20100811
  • [Presentation] Mechanism of Photosynthetic water-splitting based on the structural analysis of photosystem II2010

    • Author(s)
      Jian-Ren Shen
    • Organizer
      Inaugural Conference on Molecular & Functional Catalysis (ICMFC-1)
    • Place of Presentation
      Singapore
    • Year and Date
      20100711-20100715
  • [Presentation] 光化学系II複合体結晶の分解能向上2010

    • Author(s)
      川上恵典, 梅名泰史, 田代隆慶, 神谷信夫, 沈建仁
    • Organizer
      第10回日本蛋白質科学会年会
    • Place of Presentation
      札幌
    • Year and Date
      20100616-20100618
  • [Presentation] Mechanism of Photosynthetic Solar Energy-Conversion and Water-Splitting in Biological Systems2010

    • Author(s)
      Jian-Ren Shen
    • Organizer
      4th International Symposium on Commemoration of Exchange Agreement between National Taiwan University and Okayama
    • Place of Presentation
      Okayama, Japan
    • Year and Date
      2010-09-09
  • [Book] Red Algae in The Genomics Age2010

    • Author(s)
      Enami I., Adachi H., Shen J.-R.
    • Total Pages
      373-389
    • Publisher
      Springer, Dordrecht
  • [Book] Methods in Molecular Biology2010

    • Author(s)
      Shen J.-R., Kawakami K., Koike H.
    • Total Pages
      41-51
    • Publisher
      Humana Press, Springer NewYork Dordrecht Heidelberg London
  • [Book] Methods in Molecular Biology2010

    • Author(s)
      Yasusi Yamamoto, Jing Leng, Jian-Ren Shen
    • Total Pages
      1-10
    • Publisher
      Humana Press, Springer NewYork Dordrecht Neidelberg London

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Published: 2012-07-19  

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