天然変性タンパク質をハブとするカルビン回路高次複合体の構造解析

• Project Area: Structural basis of cell-signalling complexes mediating signal perception, transduction and responses
• Project/Area Number: 25121719
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Osaka University
• Principal Investigator: 松村 浩由 大阪大学, 工学(系)研究科(研究院), 准教授 (30324809)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥8,840,000 (Direct Cost: ¥6,800,000、Indirect Cost: ¥2,040,000); Fiscal Year 2014: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2013: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
• Keywords: カルビン回路 / 天然変性タンパク質 / 超分子複合体 / 阻害複合体 / 調節機構 / 天然変性蛋白質

Outline of Annual Research Achievements

カルビン回路高次複合体の形成によるカルビン回路酵素の協調的な阻害、ならびに天然変性蛋白質CP12の段階的折り畳み機構を解明するために、ホスホリブロキナーゼ（PRK）単体のX線結晶構造解析に続いて、PRKの反応機構を決定すべくPRK基質アナログ複合体の構造解析を行った。その結果、PRKのリン酸転移反応機構を解明できた。さらに、グリセルアルデヒド　3-リン酸デヒドロゲナーゼ(GAPDH）とCP12に、どのようにPRKが結合し、さらにCP12が連続して折りたたまれるのかを解明すべく、CP12の部位特異的変位体を5種作成し、Native-PAGEによる結合実験と表面プラズモン共鳴法を用いたキネティクス解析を行った。

Research Progress Status

26年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

26年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] The N-terminal acidic residue of the cytosolic helix 8 of an odorant receptor is responsible for different response dynamics via G-protein (2015)
  • Author(s): T. Kawasaki, T. Saka, S. Mine, E. Mizohata, T. Inoue, H. Matsumura, T. Sato
  • Journal Title: FEBS Lett. Volume: 589 Issue: 10 Pages: 1136-1143
  • DOI: 10.1016/j.febslet.2015.03.025
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] A comparative analysis of microgravity and earth grown thermostable T1 lipase crystals using HDPCG apparatus (2015)
  • Author(s): R.N. Abd Rahman, M.S. Ali, S. Sugiyama, A.T. Leow, T. Inoue, M. Basri, A.B. Salleh, and H. Matsumura
  • Journal Title: Protein Pept. Lett. Volume: 22 Pages: 173-179
  • Peer Reviewed
• [Journal Article] Crystal structure of FtsA from Staphylococcus aureus (2014)
  • Author(s): J. Fujita, Y. Maeda, C. Nagao, Y. Tsuchiya, Y. Miyazaki, M. Hirose, E. Mizohata, Y. Matsumoto, T. Inoue, K. Mizuguchi, and H. Matsumura
  • Journal Title: FEBS Lett. Volume: 588 Issue: 10 Pages: 1879-1885
  • DOI: 10.1016/j.febslet.2014.04.008
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Structural basis for the recognition-evasion arms race between Tomato mosaic virus and the resistance gene Tm-1 (2014)
  • Author(s): K. Ishibashi, Y. Kezuka, C. Kobayashi, M. Kato, T. Inoue, T. Nonaka, M. Ishikawa, H. Matsumura, and E. Katoh
  • Journal Title: Proc. Natl. Acad. Sci. USA Volume: 111 Issue: 33
  • DOI: 10.1073/pnas.1407888111
  • Peer Reviewed
• [Journal Article] Strategy for cold adaptation of the tryptophan xynthase a subunit from the psychrophile Shewanella frigidimarina K14-2: crystal structure and physicochemical properties (2014)
  • Author(s): Daisuke Mitsuya, Shun-ichi Tanaka, Hiroyoshi Matsumura, Naoto Urano, Kazufumi Takano, Kyoko Ogasahara, Michiyo Kakehira, Katsuhide Yutani, Masami Ishida
  • Journal Title: The Journal of Biochemistry Volume: 155 Issue: 2 Pages: 73-82
  • DOI: 10.1093/jb/mvt098
  • Peer Reviewed
• [Journal Article] Structural insights into the function of a thermostable copper-containing nitrite reductase (2014)
  • Author(s): Y Fukuda, et al.
  • Journal Title: The Journal of Biochemistry Volume: 155 Issue: 2 Pages: 123-135
  • DOI: 10.1093/jb/mvt107
  • Peer Reviewed
• [Journal Article] Structure of peroxiredoxin from anaerobic hyperthermophilic archaeon Pyrococcus horikoshii (2013)
  • Author(s): T. Nakamura*, A. Mori, M. Niiyama, H. Matsumura, C. Tokuyama, J. Morita, K. Uegaki and T. Inoue
  • Journal Title: Acta. Cryst. Volume: F69 Issue: 7 Pages: 719-722
  • DOI: 10.1107/s1744309113014036
  • Peer Reviewed
• [Journal Article] Expression, purification, crystallization and preliminary crystallographic study of FtsA from methicillin-resistant Staphylococcus aureus (2013)
  • Author(s): J. Fujita, Y. Miyazaki, M. Hirose, C. Nagao, E. Mizohata, Y. Matsumoto, K. Mizuguchi, T. Inoue, and H. Matsumura
  • Journal Title: Acta. Cryst. Volume: F69 Issue: 8 Pages: 895-898
  • DOI: 10.1107/s1744309113017727
  • Peer Reviewed
• [Journal Article] 転写制御因子Tup1-N 末端ドメインのX線結晶構造とその複合体形成メカニズム (2013)
  • Author(s): 中村太一、松村浩由、井上豪、日下菜花、田中直子、向由起夫
  • Journal Title: 日本結晶学会誌 Volume: 55 Pages: 110-115
  • NAID: 10031168499
  • Peer Reviewed
• [Presentation] MRSA由来細胞分裂タンパク質FtsAの構造―機能相関解析 (2014)
  • Author(s): 藤田純三、前田陽子、齋藤有紀、清水伸隆、野田勝紀、長尾知生子、土屋裕子、溝端栄一、内山進、松本佳巳、水口賢司、井上豪、松村浩由
  • Organizer: 日本結晶学会
  • Place of Presentation: 東京
  • Year and Date: 2014-11-01 – 2014-11-03
• [Presentation] MRSA由来細胞分裂タンパク質FtsAのX線結晶構造解析 (2014)
  • Author(s): 藤田純三、前田陽子、宮﨑祐満、廣瀬未果、溝端栄一、井上豪、松村浩由
  • Organizer: 日本蛋白質科学会
  • Place of Presentation: 横浜
  • Year and Date: 2014-06-25 – 2014-06-27
• [Presentation] CP12-GAPDH-PRK複合体形成によるカルビン回路調節機構の解明 (2014)
  • Author(s): 松村浩由、清水伸隆、木津奈都子、吉田将康、荒武将弘、溝端栄一、田茂井政宏、和田野晃、重岡成、井上豪
  • Organizer: 日本農芸化学会大会
  • Place of Presentation: 東京
• [Presentation] Structural basis for assembly of the CP12-GAPDH-PRK complex (2014)
  • Author(s): Hiroyoshi Matsumura, Nobutaka Shimizu, Natsuko Kizu, Masayasu Yoshida, Masahiro Aratake, Eiichi Mizohata, Masahiro Tamoi, Akira Wadano, Sshigeru Shigeoka, Tsuyoshi Inoue
  • Organizer: 日本農芸化学会2014年度大会
  • Place of Presentation: 富山
• [Presentation] カルビン回路調節システムの構造基盤 (2013)
  • Author(s): 松村浩由
  • Organizer: アグリバイオシンポジウム 2013
  • Place of Presentation: 奈良
  • Invited
• [Presentation] SAXS and XRD analyses of the Calvin cycle regulatory complexes (2013)
  • Author(s): Hiroyoshi Matsumura, Nobutaka Shimizu Tsuyoshi Inoue
  • Organizer: 日本生物物理学会年会
  • Place of Presentation: 京都
  • Invited
• [Presentation] 結晶解析と溶液散乱による相関解析 (2013)
  • Author(s): 松村 浩由
  • Organizer: タンパク質X線溶液散乱講習会
  • Place of Presentation: 筑波
  • Invited