Mechanism of leucine signaling of Thermus thermophilus

• Project/Area Number: 22780084
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Applied biochemistry
• Research Institution: The University of Tokyo
• Principal Investigator: TOMITA Takeo 東京大学, 生物生産工学研究センター, 助教 (50447364)
• Project Period (FY): 2010 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2011: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2010: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: 好熱菌 / グルタミン酸脱水素酵素 / アロステリック調節 / グローバル調節 / ロイシン / 遺伝子発現調節

Research Abstract

We revealed that glutamate dehydrogenase(GDH) from Thermus thermophilus formed hetero-hexameric structure consisted from homologous two subunits ; catalytic subunit(GdhB) and regulatory subunit(GdhB), and this complex was subject to allosteric activation by hydrophobic amino acids especially leucine. We succeeded to determine the crystal structure of GdhA/GdhB/Leu complex and revealed the allosteric mechanism of GDH. GDH2 from human(hGDH2) is also subject to allosteric activation by leucine and we found that important residues for binding of leucine were conserved in hGDH2. The mutational analyses of hGDH2 revealed that hGDH2 was also allosterically activated by leucine with a similar manner with GDH from T. thermophilus.
We also performed comprehensive analysis of gene regulation by means of DNA microarray tecnique and suggested that leucine was a signal of global regulation in T. thermophilus.

Research Products

• [Journal Article] ロイシンによるグルタミン酸脱水素酵素の活性調節機構 (2012)
  • Author(s): 富田武郎、西山真
  • Journal Title: 化学 Volume: 67巻
• [Journal Article] Structural basis for leucine-induced allosteric activation of glutamate dehydrogenase (2011)
  • Author(s): T. Tomita, T. Kuzuyama, M. Nishiyama
  • Journal Title: J. Biol. Chem. Volume: 286 Pages: 37406-13
  • Peer Reviewed
• [Journal Article] Structural basis for leucine-induced allosteric activation of glutamate de hydrogenase (2011)
  • Author(s): Tomita, T., Kuzuyama, T., Nishiyama, M.
  • Journal Title: J.Biol.Chem. Volume: 286 Pages: 37406-13
  • Peer Reviewed
• [Journal Article] Hetero-oligomeric glutamate dehydrogenase from Thermus thermophilus (2010)
  • Author(s): T. Tomita, T. Miyazaki, J. Miyazaki, T. Kuzuyama, M. Nishiyama
  • Journal Title: Microbiology Volume: 156 Pages: 3801-13
  • Peer Reviewed
• [Presentation] 高度好熱菌Thermus thermophilus由来グルタミン酸脱水素酵素のロイシンによるアロステリック調節機構の構造基盤 (2012)
  • Author(s): 富田武郎、葛山智久、西山真
  • Organizer: 2011年度日本農芸化学会大会
  • Place of Presentation: 京都
• [Presentation] Mechanism for allosteric regulation of glutamate dehydrogense from Thermus thermophilus (2012)
  • Author(s): 富田武郎、西山真
  • Organizer: International Union of Microbiological Societies 2011 Congress
  • Place of Presentation: 札幌
• [Presentation] ロイシンによるグルタミン酸脱水素酵素のアロステリック調節機構の解析 (2012)
  • Author(s): 富田武郎、葛山智久、西山真
  • Organizer: 日本農芸化学会2012年度大会
  • Place of Presentation: 京都女子大学(京都)
• [Presentation] 高度好熱菌Thermus thermophilus由来グルタミン酸脱水素酵素のロイシンによるアロステリック調節の構造基盤 (2011)
  • Author(s): 富田武郎、葛山智久、西山真
  • Organizer: 日本農芸化学会2011年度大会(京都)
  • Place of Presentation: 京都
  • Year and Date: 2011-03-27
• [Presentation] Mechanism of leucine-induced allosteric activation of glutamate dehydrogenase (2011)
  • Author(s): 富田武郎、西山真
  • Organizer: 第1回モデル生物丸ごと一匹学会
  • Place of Presentation: 兵庫
• [Presentation] Leucine-mediated signal transduction in Thermus thermophilus (2011)
  • Author(s): 松下創、富田武郎、葛山智久、西山真
  • Organizer: 第1回モデル生物丸ごと一匹学会
  • Place of Presentation: 兵庫
• [Presentation] グルタミン酸脱水素酵素の新規活性調節機構の発見 (2011)
  • Author(s): 西山真、富田武郎
  • Organizer: 2010年度日本農芸化学会大会
  • Place of Presentation: 東京
• [Presentation] 高度好熱菌Thermus thermophilusのグルタミン酸脱水素酵素(GDH)の活性調節サブユニットGdhAの結晶構造解析 (2011)
  • Author(s): 富田武郎、葛山智久、西山真
  • Organizer: 2010年度日本農芸化学会大会
  • Place of Presentation: 東京
• [Presentation] Mechanism of leucine-induced allosteric activation of glutamate dehydro genase (2011)
  • Author(s): 富田武郎、西山真
  • Organizer: 第1回モデル生物丸ごと一匹学会
  • Place of Presentation: 理化学研究所播磨研究所(兵庫)
• [Presentation] Thermus thermophilus由来グルタミン酸脱水素酵素における活性調節の分子機構の解析 (2010)
  • Author(s): 富田武郎、西山真
  • Organizer: 高度好熱菌丸ごと一匹プロジェクト第9回連携研究会
  • Place of Presentation: 播磨(Spring-8)
  • Year and Date: 2010-08-20
• [Presentation] Mechanism for Allosteric Regulation of Glutamate Dehydrogenase from Thermus thermophilius (2010)
  • Author(s): 富田武郎、葛山智久、西山真
  • Organizer: アメリカ結晶学会2010 Annual meeting
  • Place of Presentation: シカゴ USA
  • Year and Date: 2010-07-25
• [Presentation] Mulecular mechanism for allosteric regulation of glutamatedehydrogenase from Thermus thermophilus (2010)
  • Author(s): 富田武郎、西山真
  • Organizer: 高度好熱菌丸ごと一匹プロジェクト第9回連携研究会
  • Place of Presentation: 兵庫
• [Presentation] Regulatory mechanism of glutamate dehydrogenase from Thermus thermophilus (2010)
  • Author(s): 富田武郎、西山真
  • Organizer: 2010 Annual meeting of American Crystallographic Association
  • Place of Presentation: (ACA)(シカゴ)
• [Book] 化学 (2012)
  • Author(s): 富田武郎、西山真
  • Total Pages: 2
  • Publisher: 化学同人
• [Remarks]
  • URL: http://park.itc.u-tokyo.ac.jp/biotec-res-ctr/saiboukinou/