Structure and function relationships of higher plant glutamine synthetase

• Project/Area Number: 19570105
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Structural biochemistry
• Research Institution: University of Yamanashi (2008); Osaka University (2007)
• Principal Investigator: KUSUNOKI Masami University of Yamanashi, 大学院・医学工学総合研究部, 教授 (90135749)
• Co-Investigator(Kenkyū-buntansha): HASE Toshiharu 大阪大学, たんぱく質研究所, 教授 (00127276)
• Project Period (FY): 2007 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2008: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000); Fiscal Year 2007: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: X線 / 結晶 / アンモニア同化 / 植物生理学 / 構造生物学 / グルタミン合成酵素 / 立体構造 / 窒素代謝 / 酵素 / 植物

Research Abstract

植物のグルタミン合成酵素(GS)は、細胞質局在型GS1とプラスチド局在型GS2が存在し、GS1は多重遺伝子族によりコードされている。
GS1a His249Ala/MetSox-P/ADP, GS1a WT/PPT/AMPPNP の2種類の結晶解析を行った。前者から、His249の側鎖と基質の水素結合が基質認識に重要な役割を担っていること、後者からPPTのリン酸化前とリン酸化後で酵素側の構造変化が無いことも判明した。

Research Products

• [Journal Article] New Role of Flavin as a General Acid-Base Catalyst with No Redox Function in Type 2 Isopentenyl-diphosphate Isomerase (2009)
  • Author(s): Hideaki Unno
  • Journal Title: Journal of Biological Chemistry 284 Pages: 9160-9167
  • Peer Reviewed
• [Journal Article] Structural Basis for Substrate Recognition and Hydrolysis by Mouse Carnosinase CN2 (2008)
  • Author(s): Unno, H., Yamashita, H., Ujita, S., Okumura, N., Otani, H., Okumura, A., Nagai, K. and Kusunoki, M
  • Journal Title: J. Biol.Chem. 283 Pages: 27289-27299
  • Peer Reviewed
• [Journal Article] Crystallization and preliminary X-ray analysis of isomaltase from Saccharomyces cerevisiae (2008)
  • Author(s): Keizo Yamamoto
  • Journal Title: Acta Crystallographica Section F 64 Pages: 1024-1026
  • Peer Reviewed
• [Journal Article] Structural Basis for Substrate Recognition and Hydrolysis by Mouse Carnosinase CN2 (2008)
  • Author(s): Hideaki Unno
  • Journal Title: Journal of Biological Chemistr 283 Pages: 27289-27299
  • Peer Reviewed
• [Journal Article] C-Type Lectin-like Carbohydrate-Recognition of the Hemolytic Lectin CEL-III Containing Ricin-type β-Trefoil Folds (2007)
  • Author(s): Hatakeyama, T., Unno, H., Kouzuma, Y., Uchida, T., Eto, S., Hidemura, H., Kato, N., Yonekura, M. and Kusunoki, M.
  • Journal Title: J. Biol. Chem. 282 Pages: 37826-37835
  • Peer Reviewed
• [Journal Article] Structural and mutational studies of anthocyanin malonyltransferases establish the features of BAHD enzyme catalysis (2007)
  • Author(s): Unno, H., Ichimaida, F., Suzuki, H., Takahashi, S., Tanaka, Y., Saito, A., Nishino, T., Kusunoki, M. and Nakayama, T.
  • Journal Title: Journal of Biological Chemistry Vol. 282 Pages: 15182-15822
  • Peer Reviewed
• [Journal Article] Structural and mutational studies of anthocyanin malonyltransferase establish the features of BARD enzyme catalysis (2007)
  • Author(s): Hideaki Unno
  • Journal Title: Journal of Biological Chemistry 282 Pages: 15812-15822
  • Peer Reviewed
• [Journal Article] Functional, structural and spectroscopic characterization of a glutathione-ligated [2Fe-2S] cluster in poplar glutaredoxin C1 (2007)
  • Author(s): Nicolas Rouhier
  • Journal Title: Proceedings of National Academy of Sciences, USA 104 Pages: 7379-7384
  • Peer Reviewed
• [Journal Article] C-Type Lectin-like Carbohydrate-Recognition of the Hemolytic Lectin CEL-III Containing Ricin-type β-Trefoil Folds (2007)
  • Author(s): Tomomitsu Hatakeyama
  • Journal Title: Journal of Biological Chemistry 282 Pages: 37826-37835
  • Peer Reviewed
• [Journal Article] Cloning and Characterization of Ferredoxin and Ferredoxin-NADP^+ Reductase from Human Malaria Parasite (2007)
  • Author(s): Yoko Kimata-Ariga
  • Journal Title: Journal of Biochemistry 141 Pages: 421-428
  • Peer Reviewed
• [Presentation] Structural study of enzyme inhibitor compleχes of eukaryotic glutamine synthetase from Zea mays (2008)
  • Author(s): Takeshi Ozaki
  • Organizer: 国際結晶学会
  • Place of Presentation: 大阪
  • Year and Date: 2008-08-28
• [Presentation] Structural study of enzyme inhibitor complexes of eukaryotic glutamine synthetase from Zea mays (2008)
  • Author(s): Ozaki,T., Unno, H., Hase, T., Kusunoki,M.
  • Organizer: 国際結晶学会
  • Place of Presentation: 大阪
  • Year and Date: 2008-08-25
• [Presentation] Structural and Mutational Studies of Anthocyanin Malonyltransferases Establish the Features of BAHD Enzyme Catalysis (2007)
  • Author(s): 尾崎健
  • Organizer: 第8回アジア結晶学会
  • Place of Presentation: 台湾、台北市
  • Year and Date: 2007-11-05
• [Presentation] 植物グルタミン合成酵素の活性中心構造の改変による基質認識機構の解析 (2007)
  • Author(s): 山田雅子, 尾崎健, 楠木正巳, 長谷俊治
  • Organizer: 第80回日本生化学会、第30回日本分子生物学会
  • Place of Presentation: 横浜パシフィコ
• [Presentation] トウモロコシ由来のグルタミン合成酵素のH249A変異酵素の構造学的研究 (2007)
  • Author(s): 尾崎健・海野英昭・長谷俊治・楠木正巳
  • Organizer: 日本結晶学会
  • Place of Presentation: 東京工業大学大岡山キャンパス
• [Remarks]
  • URL: http://www.protein.osaka-u.ac.jp/rcsfp/pi/kusunoki/research_theme/GS.html