The coupling mechanism of glutaminase and kinase in GatCAB

• Project/Area Number: 19370037
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Structural biochemistry
• Research Institution: Hokkaido University
• Principal Investigator: YAO Min Hokkaido University, 大学院・先端生命科学研究院, 准教授 (40311518)
• Co-Investigator(Kenkyū-buntansha): TANAKA Isao 北海道大学, 大学院・先端生命科学研究院, 教授 (70093052)
• Project Period (FY): 2007 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥18,720,000 (Direct Cost: ¥14,400,000、Indirect Cost: ¥4,320,000); Fiscal Year 2008: ¥7,410,000 (Direct Cost: ¥5,700,000、Indirect Cost: ¥1,710,000); Fiscal Year 2007: ¥11,310,000 (Direct Cost: ¥8,700,000、Indirect Cost: ¥2,610,000)
• Keywords: GatCAB / グルタミナーゼ / キナーゼ / アンモニアチャネル / X線構造解析

Research Abstract

GatCABのアンモニアチャネルの開閉に重要と推定した残基の変異体調製に成功し、in vivoアッセイによって、E125が直接アンモニアの輸送に関与していることが検証された。SAXSを用いて得られた溶液中の構造情報は、tRNA^(Gln)のU1-A72がcradleドメインで、D-loopがC-tailドメインで認識することを示唆した。また、GatCABの1.9A分解能での構造解析に成功し、tRNA認識に重要であるGatBのC-tailドメインの構造を明らかにし、GatBに特徴的な配列を見い出した。更にGatCABとtRNA^(Gln)変異体のアッセイを行い、GatBのcradleドメインの3_(10)-helixとC-tailドメインに保存されたGlyがtRNA^(Gln)認識に重要であることが明らかになった。

Research Products

• [Journal Article] Structural, Functional Characterization of the LldR from Corynebacterium glutamicum : a Transcriptional Repressor Involved in l-Lactate and Sugar Utilization (2008)
  • Author(s): Yong-Gui Gao, Hiroaki Suzuki, Hiroshi Itou, Yong Zhou, Yoshikazu Tanaka, Masaaki Wachi, Nobuhisa Watanabe, Isao Tanaka, Min Yao
  • Journal Title: Nucl. Acid Res 36 Pages: 7110-7123
  • Peer Reviewed
• [Journal Article] Makoto Kimura, Crystal structure of an archaeal Ski2p-like protein from Pyrococcus horikoshii OT3 (2008)
  • Author(s): Xiaodong Zhang, Takashi Nakashima, Yoshimitsu Kakuta, Min Yao, Isao Tanaka
  • Journal Title: Protein Sci 17 Pages: 136-145
  • Peer Reviewed
• [Journal Article] Structural and Functional Characterization of the LldR from Corynebacterium glutamicum : a Transcriptional Repressor Involved in 1-Lactate and Sugar Utilization (2008)
  • Author(s): Yong-Gui Gao
  • Journal Title: Nucl. Acid Res. 36 Pages: 7110-7123
  • Peer Reviewed
• [Journal Article] Crystal structure of an archaeal Ski2p-like protein from Pyrococcus horikoshii OT3 (2008)
  • Author(s): Xiaodong Zhang
  • Journal Title: Protein Sci. 17 Pages: 136-145
  • Peer Reviewed
• [Presentation] 真正細菌型GatCABと古細菌型GatCABの比較 (2009)
  • Author(s): 中村彰良、山根潤二、Zhou Qian、姚閔、田中勲
  • Organizer: 日本生物物理学会北海道支部例会
  • Place of Presentation: 札幌市
  • Year and Date: 2009-03-09
• [Presentation] 真正細菌型GatCABのtRNA認識機構と分子進化、BMB2008 (2008)
  • Author(s): 中村彰良、山根潤二、姚閔、田中勲
  • Organizer: 第31回日本分子生物学会年会, 第81回日本生化学会大会 合同大会
  • Place of Presentation: 神戸市
• [Presentation] tRNAGln recognition loop in bacterial GatCAB, AARS2008 (International Conference on Aminoacyl-tRNA Synthetases (2008)
  • Author(s): Akiyoshi Nakamura, Junji Yamane, Min Yao, Isao Tanaka
  • Organizer: from Basic Mechanisms to Systems Biology
  • Place of Presentation: France
• [Presentation] High resolution structure of bacterial GatCAB reveals the C-tail domain structure in GatB (2008)
  • Author(s): Akiyoshi Nakamura, Jyunji Yamane, Min Yao, Isao Tanaka
  • Organizer: International Union of Crystallography (IUCr2008)
  • Place of Presentation: Osaka
• [Presentation] 真正細菌型GatCABが獲得したtRNA Gln認識ループ (2008)
  • Author(s): 中村彰良、山根潤二、姚閔、田中勲
  • Organizer: 日本分子生物学会第8回春季シンポジウム
  • Place of Presentation: 札幌市
• [Presentation] High resolution structure of bacterial GatCAB reveals the C-tail domain structure in GatB (2008)
  • Author(s): Akiyoshi Nakamura
  • Organizer: XXI Congress of the International Union of Crystallography (IUCr2008)
  • Place of Presentation: Osaka, Japan
• [Presentation] tRNAGln recognition loop in bacterial GatCAB (2008)
  • Author(s): Akiyoshi Nakamura
  • Organizer: AARS2008 (International Conference on Aminoacyl-tRNA Synthetases : from Basic Mechanisms to Systems Biology)
  • Place of Presentation: Veyrier du Lac, France
• [Presentation] 黄色ブドウ球菌由来GatCABの高分解能X線結晶構造解析 (2007)
  • Author(s): 山根潤二、中村彰良、周勇、十亀弘子、辻下英樹、武本浩、姚閔、田中勲
  • Organizer: 日本結晶学会2007年度年会
  • Place of Presentation: 東京都
• [Remarks]
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• [Remarks]
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• [Remarks]
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