Structural elucidation of the sequential signal transduction mechanism in two-component system

• Project/Area Number: 23790042
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Physical pharmacy
• Research Institution: The University of Tokyo
• Principal Investigator: UEDA Takumi 東京大学, 大学院・薬学系研究科, 助教 (20451859)
• Project Period (FY): 2011 – 2012
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2012: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2011: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: 構造生物学 / NMR / 膜蛋白質 / 転移交差飽和法 / 過渡的複合体 / ニ成分制御系 / マルチドメイン蛋白質 / 過渡的相互作用 / 膜タンパク質 / タンパク質―タンパク質相互作用 / 交差飽和法 / Protein Trans-Splicing / Segmental labeling

Research Abstract

Our NMR study based on cross-saturation method revealed that the CheA-CheY complex, which regulates bacterial chemotaxis, exists in equilibrium between catalytic and non-catalytic states, in which their catalytic sites are close to and away from each other, respectively. This binding mode allows CheA to perform two signal transduction reactions with one catalytic site.

Research Products

• [Journal Article] Efficacy of the -2-adrenergic receptor is determined byconformational equilibrium in thetransmembrane region (2012)
  • Author(s): Yutaka Kofuku, Takumi Ueda, JunyaOkude, Yutaro Shiraishi, Keita Kondo,Masahiro Maeda, Hideki Tsujishita, andIchio Shimada
  • Journal Title: Nature Commun. Volume: 3 Pages: 1045-1045
  • Peer Reviewed
• [Journal Article] Structural basis of efficient electron transport between photosynthetic membrane proteins and plastocyanin in spinach revealed using nuclear magnetic resonance (2012)
  • Author(s): Takumi Ueda, Naoko Nomoto, Masamichi Koga, Hiroki Ogasa, Yuuta Ogawa, Masahiko Matsumoto, Pavlos Stampoulis, Koji Sode, Hiroaki Terasawa, and Ichio Shimada
  • Journal Title: Plant Cell Volume: 24 Pages: 4173-4186
  • Peer Reviewed
• [Journal Article] Segmental isotopic labeling of a 140 kDa dimeric multi-domain protein CheA from Escherichia coli by expressed protein ligation and protein trans-splicing (2012)
  • Author(s): Yuichi Minato, Takumi Ueda, Asako Machiyama, Ichio Shimada, and Hideo Iwai
  • Journal Title: J.Biomol. NMR Volume: 53 Pages: 191-207
  • Peer Reviewed
• [Journal Article] Atypical membrane-embedded phosphatidylinositol3,4-bisphosphate (PI(3,4)P2)-binding site on p47phox Phox homology (PX) domain revealed by NMR. (2012)
  • Author(s): Pavlos Stampoulis, Takumi Ueda, Masahiko Matsumoto, Hiroaki Terasawa, and Ichio Shimada
  • Journal Title: J. Biol. Chem. Volume: 287 Pages: 17848-17859
  • Peer Reviewed
• [Journal Article] Functional Equilibrium of the KcsA Structure Revealed by NMR (2012)
  • Author(s): Imai S., Osawa M., Mita K., Toyonaga S., Machiyama A., Ueda T., Takeuchi K., Oiki S., and Ichio Shimada
  • Journal Title: J. Biol. Chem. Volume: 287 Pages: 39634-39641
  • Peer Reviewed
• [Journal Article] Functional dynamics of proteins revealed by NMR (2012)
  • Author(s): Masanori Osawa, Koh Takeuchi, Takumi Ueda, Noritaka Nishida, and Ichio Shimada
  • Journal Title: Curr. Opin. Struct. Biol. Volume: 22 Pages: 660-669
  • Peer Reviewed
• [Journal Article] Efficacy of the b2-adrenergic receptor is determined by conformational equilibrium in the transmembrane region (2012)
  • Author(s): Yutaka Kofuku
  • Journal Title: Nature Communicatinos Volume: 3 Pages: 1045-1045
  • Peer Reviewed
• [Journal Article] Structural Basis of Efficient Electron Transport between Photosyn thetic Membrane Proteins and Plastocyanin in Spinach Revealed using Nuclear Magnetic Resonance (2012)
  • Author(s): Takumi Ueda, Naoko Nomoto, Masamichi Koga, Hiroki Ogasa, Yuuta Ogawa, Masahiko Matsumoto, Pavlos Stampoulis, Koji Sode, Hiroaki Terasawa and Ichio Shimada
  • Journal Title: Plant Cell Volume: 24 Issue: 10 Pages: 4173-4186
  • DOI: 10.1105/tpc.112.102517
  • Peer Reviewed
• [Journal Article] Segmental isotopic labeling of a 140 kDadimeric multi-domain protein CheA from Escherichia Coli by Expressed Protein Ligation and Protein Trans-Splicing (2012)
  • Author(s): Yuichi Minato, Takumi Ueda, Asako Machiyama, Ichio Shimada, Hideo Iwai
  • Journal Title: Journal of Biomolecular NMR Volume: in press
  • Peer Reviewed
• [Journal Article] Atypical membrane-embedded p47^<phox> PI(3,4)P_2 binding site on PX domain revealed by NMR (2012)
  • Author(s): Stampoulis P, Ueda T, Matsumoto M, Terasawa H, Miyano K, Sumimoto H, and Shimada I
  • Journal Title: J. Biol. Chem Volume: 287 Issue: 21 Pages: 17848-17859
  • DOI: 10.1074/jbc.m111.332874
  • Peer Reviewed
• [Journal Article] The Atmospheric Scanning Electron Microscope with open sample space observes dynamic phenomena in liquid or gas (2011)
  • Author(s): Mitsuo Suga, Hidetoshi Nishiyama, Yuji Konyuba, Shinnosuke Iwamatsu, Yoshiyuki Watanabe, Chie Yoshiura, Takumi Ueda, andChikara Sato
  • Journal Title: Ultramicroscopy Volume: 111 Pages: 1650-1658
• [Presentation] Structural Elucidation of the AlternateSignal Transduction Mechanism of CheA (2012)
  • Author(s): Yuichi Minato, Takumi Ueda, AsakoMachiyama, Hideo Iwai, Ichio Shimada,
  • Organizer: International Conference on MagneticResonance in Biological Systems
  • Place of Presentation: リヨン(フランス)
• [Presentation] Development of interferogram extrapolation method for NMR experiments that requires accurate peakheight (2012)
  • Author(s): Takumi Ueda, Masahiko Matsumoto, Ichio Shimada
  • Organizer: ISNMR2011
  • Place of Presentation: 横浜
• [Presentation] Structural Elucidation of the Alternate Signal Transduction Mechanism of CheA and CheY (2012)
  • Author(s): Yuichi Minato
  • Organizer: International Conference on Magnetic Resonance in Biological Systems
  • Place of Presentation: リヨン, フランス
  • Invited
• [Presentation] Development of interferogram extrapolation method for NMR experiments that requires accurate peak height (2011)
  • Author(s): Takumi Ueda, Masahiko Matsumoto, Ichio Shimada
  • Organizer: ISNMR2011
  • Place of Presentation: 横浜