Crystallographic and Molecular Studies on Yeast 26S Proteasome

• Project/Area Number: 19570103
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Structural biochemistry
• Research Institution: Kyoto University
• Principal Investigator: MORIMOTO Yukio Kyoto University, 原子炉実験所, 教授 (80200450)
• Research Collaborator: NISHIO Kazuya 京都大学, 原子炉実験所, 特定助教 ; SUGIYAMA Masaaki 京都大学, 原子炉実験所, 准教授
• Project Period (FY): 2007 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2008: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2007: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
• Keywords: X線結晶構造解析 / プロテアソーム / 結晶構造解析

Research Abstract

本研究課題では(1)26Sプロテアソームの精製と結晶化(2)溶液状態での両端因子の会合、解離過程を追跡すること、を目的とした。発現系3種の酵母株のうち676株からの微結晶を得ることに成功し低分解能(35Å)ながら回折斑点を得ることができた。これとともに脱ユビキンチン化酵素(UCH37)の結晶構造解析を進め、20S本体を形成しているαサブユニットリングの形成過程、などの形態解析を行った。

Research Products

• [Journal Article] K.Mori and T.Fukunaga. SANS Simulation of Aggregated Protein in Aqueous Solution Nuclear Inst (2009)
  • Author(s): M.Sugiyama, K.Hamada, K.Kato, E.Kurimoto, K.Okamoto, Y.Morimoto, S.Ikeda, S.Naito, M.Furusaka, K.Itoh, K.Mori and T.Fukunaga
  • Journal Title: And Methods in Physic Research A 600 Pages: 272-274
  • Peer Reviewed
• [Journal Article] Structural evolution of human recombinant alphaB-crystallin under UV irradiation (2008)
  • Author(s): M.Sugiyama,, N.Fujii, Y.Morimoto, S.Kurabayashi, M.Vigild, T.Sato, T.Nakazawa, K.Itoh, K.Mori and T.Fukunaga
  • Journal Title: Biomacromolecules 9 Pages: 431-434
  • Peer Reviewed
• [Journal Article] An Abnormal pKa value of internal histidine of the insulin molecule revealed by neutron crystallographic analysis (2008)
  • Author(s): T.Ishikawa, T.Chatake, Y.Morimoto, M.Maeda, K.Kurihara, I.Tanaka and N.Niimura
  • Journal Title: BBRC 376 Pages: 32-35
  • Peer Reviewed
• [Journal Article] Preliminary time-of-flight neutron diffraction study of human deoxyhemoglobin (2008)
  • Author(s): A.Y.Kovalevsky, T.Chatake, N.Shibayama, S.-Y. Park, T.Ishikawa, M.Mustyakimov, S.Z.Fisher, P.Langan and Y.Morimoto
  • Journal Title: Acta.Cryst F64 Pages: 270-273
  • Peer Reviewed
• [Journal Article] The crystallographic study of the Deubiquitinating Enzyme UCH37 N-terminal Domain (2008)
  • Author(s): K.Nishio, K.Kawai, S.Kim, T.Mizushima, T.Yamane, J.Hamazaki, S.Murata, K.Tanaka and Y.Morimoto
  • Journal Title: KURRI Prog.Rep. 2007 P216
  • Peer Reviewed
• [Journal Article] The crystallographic study of the deubiquitinating enzyme UCH37 N-terminal domain (2008)
  • Author(s): K. Nishio, K. Kawai, S. -W. Kim, T. Mizushima, T. Yamane, J. Hamasaki, S. Murata, K. Tanaka, Y. Morimoto
  • Journal Title: Acta Cryst. A64
  • Peer Reviewed
• [Journal Article] Structural evolution of human recombinant alphaB-crystallin under UV irradiation (2008)
  • Author(s): M. Sugiyama, N. Fujii, Y. Morimoto, S. Kurabayashi, M. Vigild, T. Sato,
  • Journal Title: Biomacromolecules 9 Pages: 431-434
  • Peer Reviewed
• [Journal Article] Shape of alpha-Crystallin Analyzed by Small-Angle Neutron Scattering (2007)
  • Author(s): M. Sugiyama, N. Fujii, Y. Morimoto, T. Otomo, S. Tanaka, M. Misawa,
  • Journal Title: J. Appl. Cryst. 40
  • Peer Reviewed
• [Journal Article] 放射光・中性子結晶解析によるタンパク質の構造機能研究 (2007)
  • Author(s): 森本幸生
  • Journal Title: 波紋 17 Pages: 44-47
  • NAID: 130001998393
  • Peer Reviewed
• [Presentation] 哺乳類タンパク複合体に見られるサブユニット交換と立体構造の特徴 (2009)
  • Author(s): 森本幸生
  • Organizer: 第34回生命の起原および進化学会
  • Place of Presentation: 熊取
  • Year and Date: 2009-03-18
• [Presentation] The crystallographic study of the deubiquitinating enzyme UCH37 N-terminal domain (2008)
  • Author(s): K.Nishio, K.Kawai, S.W.Kim, T. Mizushima, T. Yamane, J. Hamasaki, S. Murata, K. Tanaka and Y. Morimoto
  • Organizer: 国際結晶学連合会
  • Place of Presentation: Osaka
  • Year and Date: 2008-08-26
• [Presentation] 脱ユビキチン化酵素UCH37N末端ドメインの結晶学的研究 (2008)
  • Author(s): 西尾和也、河合健太郎、金相佑、水島恒裕、山根隆、濱崎純、村田茂穂、田中啓二、森本幸生
  • Organizer: 日本蛋白質科学会
  • Place of Presentation: 東京
  • Year and Date: 2008-06-11
• [Presentation] SANS study of Proteasome Activator 28 in an aqueous solution. (2007)
  • Author(s): M. Sugiyama, Y. Morimoto, K. Itoh, K. Mori, T. Fukunaga, K. Hamada,
  • Organizer: Neutron in Biology 2007
  • Place of Presentation: Rutherford Appleton Laboratory, UK
  • Year and Date: 2007-07-12
• [Presentation] SANS study of Proteasome Activator 28 in an aqueous solution (2007)
  • Author(s): M.Sugiyama, Y.Morimoto, K.Itoh, K.Mori, T.Fukunaga, K.Hamada, T.Sato, H.Sahashi, E.Sakata and K.Kato
  • Organizer: Neutron in Biology
  • Place of Presentation: Oxford
  • Year and Date: 2007-07-10
• [Remarks]
  • URL: http://www.rri.kyoto-u.ac.jp/morimotolabo.html