Time-resolved structure analysis of type-II restriction endonuclease HindIII

• Project/Area Number: 22370036
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Structural biochemistry
• Research Institution: Nagoya University
• Principal Investigator: WATANABE Nobuhisa 名古屋大学, シンクロトロン光研究センター, 教授 (70212321)
• Project Period (FY): 2010 – 2012
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥18,720,000 (Direct Cost: ¥14,400,000、Indirect Cost: ¥4,320,000); Fiscal Year 2012: ¥6,370,000 (Direct Cost: ¥4,900,000、Indirect Cost: ¥1,470,000); Fiscal Year 2011: ¥6,240,000 (Direct Cost: ¥4,800,000、Indirect Cost: ¥1,440,000); Fiscal Year 2010: ¥6,110,000 (Direct Cost: ¥4,700,000、Indirect Cost: ¥1,410,000)
• Keywords: X線 / 結晶構造解析 / 制限酵素 / 反応機構 / 時間分割 / フリーズトラップ / HindIII / フリーズトラップ法 / Ｘ線 / 結晶構造 / 蛋白質 / 生物物理 / フラッシュフリーズ法

Research Abstract

We have performed the time-resolved X-ray crystal structure analysis of a type-II restriction endonuclease HindIII using the freeze trap method. Crystals of HindIII and its cognate DNA complex were soaked in a solution containing metal ion, and flash-freezed to stop its reaction after the elapse of a certain period of time. In the reaction, two metal ions are necessary, and the scission reaction takes place after binding of the second metal ion.

Research Products

• [Journal Article] Analysis of the HindIII-catalyzed reaction by time-resolved crystallography (2015)
  • Author(s): T. Kawamura, T. Kobayashi & N. Watanabe
  • Journal Title: Acta Crystallographica, section D Volume: 71 Issue: 2 Pages: 256-265
  • DOI: 10.1107/s1399004714025188
  • Peer Reviewed
• [Presentation] Time-resolved structure analysis of restriction endonuclease Hind III (2013)
  • Author(s): Kawamura, T., Kobayashi, T. and Watanabe, N
  • Organizer: International Conference on Structural Genomics 2013 - Structural Life Science
  • Place of Presentation: Keio Plaza Hotel Sapporo
• [Presentation] Time-resolved structure analysis of restriction endonuclease Hind III (2013)
  • Author(s): Kawamura, T., Kobayashi, T. and Watanabe, N.
  • Organizer: International Conference on Structural Genomics 2013 - Structural Life Science
  • Place of Presentation: 札幌市 京王プラザホテル
• [Presentation] II 型制限酵素 Hind III の反応メカニズムと 2価金属イオン (2012)
  • Author(s): 小林朋希,河村高志,渡邉信久
  • Organizer: 日本結晶学会年会
  • Place of Presentation: 東北大学片平キャンパス
  • Year and Date: 2012-10-25
• [Presentation] II型制限酵素Hind IIIの反応メカニズムと2価金属イオン (2012)
  • Author(s): 小林朋希，河村高志，渡邉信久
  • Organizer: 日本結晶学会年会
  • Place of Presentation: 東北大学 片平キャンパス
• [Presentation] Hind III-cognate DNA 複合体の新規結晶化条件とフリーズトラップ法のための装置開発 (2011)
  • Author(s): 小林朋希,河村高志,渡邉信久
  • Organizer: 日本蛋白質科学会年会
  • Place of Presentation: ホテル阪急エクスポパーク
• [Presentation] Hind III-cognate DNA複合体の新規結晶化条件とフリーズトラップ法のための装置開発 (2011)
  • Author(s): 小林朋希，河村高志，渡邉信久
  • Organizer: 日本蛋白質科学会年会
  • Place of Presentation: ホテル阪急エクスポパーク
• [Presentation] フリーズトラップ用時分割フラッシュクーリング結晶マウント装置の開発 (2010)
  • Author(s): 渡邉信久,小林朋希,河村高志
  • Organizer: 日本結晶学会年会
  • Place of Presentation: 大阪大学コンベンションセンター
  • Year and Date: 2010-12-05