Method for crystal quality improvement of protein crystal

2013 Fiscal Year Final Research Report

• Project/Area Number: 24657079
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Single-year Grants
• Research Field: Structural biochemistry
• Research Institution: High Energy Accelerator Research Organization
• Principal Investigator: SENDA Toshiya 大学共同利用機関法人高エネルギー加速器研究機構, 物質構造科学研究所, 教授 (30272868)
• Project Period (FY): 2012-04-01 – 2014-03-31
• Keywords: 結晶工学 / クライオプロテクタント / タンパク質 / 結晶化 / 分解能 / X線結晶構造解析 / 回折データ測定

Research Abstract

The multi-step soaking method is a novel approach to the post-crystallization treatment used to improve quality of protein crystals. This new method improved the crystal quality of CagA, a type IV secretion effector derived from Helicobacter pylori, from 10 angstrom to 3.1 angstrom resolution. In addition, this method were applied to the other protein, and improved its crystal quality from 3.5 angstrom resolution to 2.1 angstrom resolution.

Research Products

• [Journal Article] Structural basis of theγ-lactone-ring formation in ascorbic acid biosynthesis by the senescence marker protein-30/gluconolactonase (2013)
  • Author(s): S. Aizawa, M. Senda, A. Harada, N. Maruyama, T. Ishida, T. Aigaki, A. Ishigami, T. Senda
  • Journal Title: PLoS One Volume: 8 (1) Pages: e53706
  • DOI: 10.1371/journal.pone.0053706
  • Peer Reviewed
• [Journal Article] Tertiary structure-function analysis reveals the pathogenic signaling potentiation mechanism of Helicobacter pylori oncogenic effector CagA (2013)
  • Author(s): T. Hayashi, M.Senda, H. Morohashi, H. Higashi, M. Horio, L. Nagase, D.Sasaya, T. Shimizu, N. Venugopalan, H. Kumeta, N. Noda, F. Inagaki, T. Senda, M. Hatakeyama
  • Journal Title: Cell Host Microbe Volume: 19 Pages: 20-33
  • DOI: 10.1016/j.chom.2012.05.010
  • Peer Reviewed
• [Journal Article] Crystallization and preliminary crystallographic analysis of D-aspartate oxidase from porcine kidney (2012)
  • Author(s): M . Senda, A. Yamamoto, H. Tanaka, T. Ishida, K. Horiike, T. Senda
  • Journal Title: Acta Crystallogr Sect F Volume: 68 Pages: 644-646
  • DOI: 10.1107/S1744309112013243
  • Peer Reviewed
• [Presentation] Crystal quality improvement with cryoprotectants (2013)
  • Author(s): Toshiya Senda
  • Organizer: International Conference on Structural Genomics 2013 -Structural Life Science
  • Place of Presentation: 京王プラザホテル札幌(札幌)
  • Year and Date: 2013-07-30
• [Presentation] A novel approach of crystal-quality improvement by"the multi-step soaking method" (2013)
  • Author(s): Miki Senda
  • Organizer: American Crystallographic Association Annual Meeting 2013
  • Place of Presentation: シェラトンワイキキホテル(ハワイ)
  • Year and Date: 2013-07-24
• [Presentation] 「multi-step soaking method」によるピロリ菌の発がんタンパク質CagA結晶の改善 (2013)
  • Author(s): 千田美紀
  • Organizer: 第13回日本蛋白質科学会年会
  • Place of Presentation: とりぎん文化会館(鳥取)
  • Year and Date: 2013-06-12
• [Presentation] クライオプロテクタントを用いた結晶工学的処理によるHelicobacter pylori由来CagA結晶の改善 (2012)
  • Author(s): 千田美紀
  • Organizer: 大阪大学蛋白質研究所セミナー
  • Place of Presentation: 大阪大学(大阪)
  • Year and Date: 2012-09-21
• [Presentation] Improve the efficiency and reproducibility of protein crystallization (2012)
  • Author(s): Miki Senda
  • Organizer: American Crystallographic Association Annual Meeting 2012
  • Place of Presentation: The Westin Boston Waterfront hotel(ボストン)
  • Year and Date: 2012-07-31
• [Presentation] 「すぐ見る法」による蛋白質の結晶化〜スクリーニングの効率化と再現性の向上に向けて (2012)
  • Author(s): 千田美紀
  • Organizer: 第12回日本蛋白質科学会年会
  • Place of Presentation: 名古屋大学(名古屋)
  • Year and Date: 2012-06-20
• [Book] 実験医学増刊構造生命科学で何がわかるのか、何ができるのか (2014)
  • Author(s): 千田俊哉, 他
  • Total Pages: 193
  • Publisher: 羊土社