Structural fluctuations and their role in the folding-assisting function of the chaperonin complex

2010 Fiscal Year Self-evaluation Report

• Project Area: Molecular Science of Fluctuations toward Biological Functions
• Project/Area Number: 20107009
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Science and Engineering
• Research Institution: National Institutes of Natural Sciences Okazaki Research Facilities
• Principal Investigator: KUWAJIMA Kunihiro National Institutes of Natural Sciences Okazaki Research Facilities, 岡崎統合バイオサイエンスセンター, 教授 (70091444)
• Project Period (FY): 2008 – 2012
• Keywords: 分子機械 / 分子認識 / 蛋白質 / アロステリック転移 / シャペロニン

Research Abstract

分子シャペロンは細胞内の蛋白質フォールディングに関わり,分子レベルの生命現象である蛋白質フォールディングと細胞レベルの生命現象とを結びつける重要な概念である。本研究では,分子シャペロンの構造揺らぎと機能発現との関係を物理化学的に明らかにする。そのため,ジメチルスルフォキシド(DMSO)停止水素重水素(H/D)交換法を蛋白質超分子であるシャペロニン複合体のH/D交換反応の追跡に適用する。シャペロニン複合体中のGroES部分(分子量10,000)のH/D交換反応をから着手し,さらにインテインによる蛋白質ライゲーションを利用してGroEL部分(分子量57,000)のH/D交換解析も視野に入れる。DMSO停止H/D交換法と二次元NMRを利用して,さまざまなシャペロニン複合体中のGroESのH/D交換反応を追跡し,生物機能が構造揺らぎをどのように利用しているかを明らかにする。

Research Products

• [Journal Article] Dissecting a bimolecular process of MgATP^2- binding to the chaperonin GroEL. (2011)
  • Author(s): Chen, J., Makabe, K., Nakamura, T., Inobe, T., Kuwajima, K.
  • Journal Title: J.Mol.Biol. in the press.
  • Peer Reviewed
• [Journal Article] Different folding pathways taken by highly homologous proteins, goat α-lactalbumin and canine milk lysozyme. (2010)
  • Author(s): Nakamura, T., Makabe, K., Tomoyori, K., Maki, K., Mukaiyama, A., Kuwajima, K.
  • Journal Title: J.Mol.Biol. 396 Pages: 1361-1378
  • Peer Reviewed
• [Journal Article] Non-native α-helix formation is not necessary for folding of lipocalin : Comparison of burst-phase folding between tear lipocalin and β-lactoglobulin. (2009)
  • Author(s): Tsukamoto, S., Yamashita, T., Yamada, Y., Fujiwara, K., Maki, K., Kuwajima, K., Matsumura, Y., Kihara, H., Tsuge, H., Ikeguchi, M.
  • Journal Title: Proteins 76 Pages: 226-236
  • Peer Reviewed
• [Journal Article] Asymmetry of the GroEL-GroES complex under physiological conditions as revealed by small-angle X-ray scattering. (2008)
  • Author(s): Inobe, T., Takahashi, K., Maki, K., Enoki, S., Kamagata, K., Kadooka, A., Arai, M., Kuwajima, K.
  • Journal Title: Biophys.J. 94 Pages: 1392-1402
  • Peer Reviewed
• [Journal Article] Probing force- induced unfolding intermediates of a single staphylococcal nuclease molecule and the effect of ligand binding. (2008)
  • Author(s): Ishii, T., Murayama, Y., Katano, A., Maki, K., Kuwajima, K., Sano, M.
  • Journal Title: Biochem.Biophys.Res.Commun. 375 Pages: 586-591
  • Peer Reviewed
• [Presentation] Molecular mechanisms of the Escherichia coli chanperonin function (2011)
  • Author(s): K.Kuwajima
  • Organizer: 3rd APPA Conference
  • Place of Presentation: Shanghai University,Shanghai,China
  • Year and Date: 20110506-20110507
• [Presentation] Identification of fatty-acid binding site in the anti-tumor complex of of α-lactalbuin by 920-MHz NMR spectroscopy (2010)
  • Author(s): K.Kuwajima
  • Organizer: The 10th KIAS Conference on Protein Structure and Function
  • Place of Presentation: Korea Institute for Advanced Study(KIAS)Seoul,Korea
  • Year and Date: 20100930-20101002
• [Presentation] Is the Folding Pathway Conserved in Homologous Proteins (2010)
  • Author(s): K.Kuwajima
  • Organizer: Bit Life Science's 3rd Annual Protein and Peptide Conference After a Solution of the Machines of Life
  • Place of Presentation: Beijing International Convention Center,Beijing,China
  • Year and Date: 20100521-20100523
• [Presentation] Molecular mechanisms of protein folding (2010)
  • Author(s): K.Kuwajima
  • Organizer: 2010 Annual Meeting of Asian CORE Program Frontiers of Materials,Photo-,and Theoretical Molecular Sciences Insitute of Atomic and Molecular Sciences
  • Place of Presentation: Academia Sinica,Taipei,Taiwan
  • Year and Date: 20100228-20100302
• [Book] Water and Biomolecules-Physical Chemistry of Life Phenomena.(Kuwajima.K, Goto.Y, Hirata.F, Kataoka.M., Terazima.M, Eds.)(A.Experimental and Simulation Studies of the Folding/Unfolding of Goat α-Lactalbumin.) (2009)
  • Author(s): Kuwajima, K., Oroguchi, T., Nakamura, T., Ikeguchi, M., Kidera
  • Total Pages: 13-35
  • Publisher: Springer-Verlag.Berlin Heiderberg