Segmental labeling of the intrinsically disordered protein PQBP1 and elucidation of the molecular recognition mechanism by NMR

• Project/Area Number: 21790034
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Physical pharmacy
• Research Institution: University of Toyama
• Principal Investigator: MIZUGUCHI Mineyuki 富山大学, 大学院・医学薬学研究部(薬学), 教授 (30332662)
• Project Period (FY): 2009 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2011: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2010: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2009: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: 生体分子 / 蛋白質 / 分子認識 / NMR / タンパク質 / 天然変性蛋白質

Research Abstract

We successfully prepared segmentally labeled PQBP1, in which the C-terminal domain is 15N-labeled. Two fragments of PQBP1 were highly expressed in E. coli with the cold-shock vector, and ligated by the expressed protein ligation method. Our results will be applicable to the segmental labeling of other intrinsically disordered proteins.

Research Products

• [Journal Article] Hydrogen-bondnetwork and pH sensitivity in transthyretin : neutron crystal structure of humantransthyretin (2012)
  • Author(s): Yokoyama, T., Mizuguchi, M., Nabeshima, Y., Kusaka, K., Yamada, T., Hosoya, T., Ohhara, T., Kurihara, K., Tomoyori, K., Tanaka, I. and Niimura. N.
  • Journal Title: J Struct Biol Volume: 177 Pages: 283-290
  • Peer Reviewed
• [Journal Article] Hydrogen-bond network and pH sensitivity in transthyretin: neutron crystal structure of human transthyretin (2012)
  • Author(s): Yokoyama, T., et al.
  • Journal Title: Journal of Structural Biology Volume: 177 Pages: 283-290
  • Peer Reviewed
• [Journal Article] 天然変性タンパク質PQBP-1の揺らぎと生体機能.メディカルバイオ (2010)
  • Author(s): 水口峰之, 岡澤均
  • Journal Title: 10月号別冊「揺らぎと生体機能」 Pages: 44-48
• [Journal Article] Polyglutamine tract-bindingprotein-1 binds to U5-15kD via a continuous23-residue segment of the C-terminaldomain (2010)
  • Author(s): Takahashi, M., Mizuguchi, M., Shinoda, H., Aizawa, T., Demura, M., Okazawa H. andKawano, K
  • Journal Title: Biochim Biophys Acta Volume: 1804 Pages: 1500-1507
  • Peer Reviewed
• [Journal Article] Polyglutamine tract-binding protein-1 binds to U5-15kD via a continuous 23-residue segment of the C-terminal domain. (2010)
  • Author(s): Takahashi, M., et al.
  • Journal Title: Biochimica et Biophysica Acta-Proteins and Proteomics Volume: 1804 Pages: 1500-1507
  • Peer Reviewed
• [Journal Article] 天然変性タンパク質PQBP-1の揺らぎと生体機能 (2010)
  • Author(s): 水口峰之, 岡澤均
  • Journal Title: メディカルバイオ Volume: 10月号別冊 Pages: 44-48
• [Journal Article] A novelβ-defensin structure : big defensin changes its N-terminal structureto associate with the target membrane (2009)
  • Author(s): Kouno, T., Mizuguchi, M., Aizawa, T., Shinoda, H., Demura, M., Kawabata, S. andKawano, K
  • Journal Title: Biochemistry 48 Pages: 7629-7635
  • Peer Reviewed
• [Journal Article] Polyglutamine tract bindingprotein-1 is an intrinsically unstructuredprotein (2009)
  • Author(s): Takahashi, M., Mizuguchi, M., Shinoda, H., Aizawa, T., Demura, M., Okazawa, H. andKawano, K
  • Journal Title: Biochim Biophys Acta Volume: 1794 Pages: 936-943
  • Peer Reviewed
• [Journal Article] Polyglutamine tract binding protein-1 in an intrinsically unstructured protein. (2009)
  • Author(s): Takahashi, M., et al.
  • Journal Title: Biochimica et Biophysica Acta-Proteins and Proteomics 1794 Pages: 936-943
  • Peer Reviewed
• [Journal Article] A novel β-defensin structure : big defensin changes its N-terminal structure to associate with the target membrane. (2009)
  • Author(s): Kouno, T., et al.
  • Journal Title: Biochemistry 48 Pages: 7629-7635
  • Peer Reviewed
• [Presentation] ポリグルタミン結合タンパク質の構造生物学的研究 (2012)
  • Author(s): 水口峰之
  • Organizer: 日本薬学会第132年会
  • Place of Presentation: 札幌
  • Year and Date: 2012-03-29
• [Presentation] ポリグルタミン結合タンパク質の構造生物学的研究 (2012)
  • Author(s): 水口峰之
  • Organizer: 日本薬学会第132年会
  • Place of Presentation: 北海道大学(招待講演)
  • Year and Date: 2012-03-29
• [Presentation] Structural changes of theintrinsically disordered protein PQBP-1 (2012)
  • Author(s): Mizuguchi, M
  • Organizer: 9thJapan-Korea Bilateral Symposium onBiological NMR
  • Place of Presentation: Sapporo
  • Year and Date: 2012-03-16
• [Presentation] Structural changes of the intrinsically disordered protein PQBP-1 (2012)
  • Author(s): Mizuguchi M.
  • Organizer: 9th Japan-Korea Bilateral Symposium on Biological NMR
  • Place of Presentation: Hokkaido University(招待講演)
  • Year and Date: 2012-03-16
• [Presentation] Dynamic structural changesof the intrinsically disordered proteinPQBP-1 (2012)
  • Author(s): Mizuguchi, M
  • Organizer: 4th Japan-Korea Seminar onBiomolecular Sciences, Experiments andSimulations
  • Place of Presentation: Nara
  • Year and Date: 2012-01-11
• [Presentation] Dynamic structural changes of the intrinsically disordered protein PQBP-1 (2012)
  • Author(s): Mizuguchi M.
  • Organizer: 4th Japan-Korea Seminar on Biomolecular Sciences, Experiments and Simulations
  • Place of Presentation: Cultural Center of Todaiji-Temple(招待講演)
  • Year and Date: 2012-01-11
• [Presentation] 精神遅滞の原因タンパク質PQBP-1の構造と機能 (2011)
  • Author(s): 水口峰之
  • Organizer: 第21回WSフォーラムタンパク質・ペプチド研究の現状と展望
  • Place of Presentation: 福岡
  • Year and Date: 2011-11-19
• [Presentation] 精神遅滞の原因タンパク質PQBP-1の構造と機能 (2011)
  • Author(s): 水口峰之
  • Organizer: 第21回WSフォーラムタンパク質・ペプチド研究の現状と展望
  • Place of Presentation: 九州大学(招待講演)
  • Year and Date: 2011-11-19
• [Presentation] 天然変性蛋白質PQBP-1の動的な構造変化 (2011)
  • Author(s): 水口峰之
  • Organizer: 物理学会2011年秋季大会
  • Place of Presentation: 富山
  • Year and Date: 2011-09-22
• [Presentation] 天然変性蛋白質PQBP-1の動的な構造変化 (2011)
  • Author(s): 水口峰之
  • Organizer: 物理学会2011年秋季大会
  • Place of Presentation: 富山大学(招待講演)
  • Year and Date: 2011-09-22
• [Presentation] Fluctuation and function ofpolyglutamine tract binding protein-1 (2009)
  • Author(s): Mizuguchi, M., Takahashi, M., Okazawa, H. and Kawano, K
  • Organizer: The3rd International Symposium on MolecularScience of Fluctuations toward BiologicalFunctions
  • Place of Presentation: Nagoya
  • Year and Date: 2009-12-21
• [Presentation] Fluctuation and function of polyglutamine tract binding protein-1 (2009)
  • Author(s): Mizuguchi, M., Takahashi, M., Okazawa, H., Kawano, K.
  • Organizer: The 3rd International Symposium on Molecular Science of Fluctuations toward Biological Functions
  • Place of Presentation: Nagoya University
  • Year and Date: 2009-12-21
• [Remarks]
  • URL: http://www.pha.u-toyama.ac.jp/phypha2/index-j.html