X-ray crystallographic analysis of microsomal prostaglandin E2synthase1

• Project/Area Number: 20370042
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Structural biochemistry
• Research Institution: The Institute of Physical and Chemical Research
• Principal Investigator: AGO Hideo The Institute of Physical and Chemical Research, 宮野構造生物物理研究室, 専任研究員 (70360477)
• Project Period (FY): 2008 – 2010
• Project Status: Completed (Fiscal Year 2010)
• Budget Amount: ¥14,300,000 (Direct Cost: ¥11,000,000、Indirect Cost: ¥3,300,000); Fiscal Year 2010: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000); Fiscal Year 2009: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2008: ¥7,020,000 (Direct Cost: ¥5,400,000、Indirect Cost: ¥1,620,000)
• Keywords: アラキドン酸カスケード / 膜タンパク質 / X線結晶構造解析 / プロスタグランジンE2 / 膜蛋白質 / 結晶化 / 膜蛋白 / X線結晶構造 / 蛋白質改変 / 酵素活性 / モノクローナル抗体

Research Abstract

Human membrane protein microsomal prostaglandin E2 synthase 1(mPGES1) was overexpressed using Schizosaccharomyces pombe. The purification method was established. The recombinant mPGES1 was free from the post-translational modification except for the deletion of the N-terminal methionine residue, and had a higher specific activity. The recombinant mPGES1 was crystallized in the two crystallization conditions.

Research Products

• [Journal Article] The catalytic architecture of Leukotriene C4 synthase with two arginine residues (2011)
  • Author(s): Saino H., Ago H., et al.
  • Journal Title: J.Biol.Chem. (In press)
  • Peer Reviewed
• [Journal Article] システインロイコトリエン研究の新知見 (2010)
  • Author(s): 吾郷日出夫、宮野雅司
  • Journal Title: 日本結晶学会誌 52 Pages: 69-75
  • NAID: 10026107914
• [Presentation] プロスタグランジンE_2合成酵素1の結晶化-結晶の改善 (2010)
  • Author(s): 齊野廣道
  • Organizer: 日本結晶学会平成22年度年会
  • Place of Presentation: 吹田市
  • Year and Date: 2010-12-05
• [Presentation] プロスタグランジンE_2合成酵素1の結晶化法の改良 (2010)
  • Author(s): 齊野廣道
  • Organizer: 平成22年度日本結晶学会年会
  • Place of Presentation: 吹田市
  • Year and Date: 2010-12-05
• [Presentation] Synchrotron radiation structural biology of proteins functioning at biological memebrane (2010)
  • Author(s): 吾郷日出夫
  • Organizer: 第27回内藤コンファレンス 生体膜ダイナミクスと脂質生物学[I]
  • Place of Presentation: 札幌市
  • Year and Date: 2010-06-30
• [Presentation] Synchrotron radiation structural biology of proteins functioning at biological membrane (2010)
  • Author(s): 吾郷日出夫
  • Organizer: 第27回内藤コンファレンス生体膜ダイナミクスと脂質生物学[I]
  • Place of Presentation: 札幌市
  • Year and Date: 2010-06-30
• [Presentation] The distinct role of two arginine residues in the leukotriene A4-specific glutathione S-transferase activity of LTC4 synthase (2010)
  • Author(s): 齊野廣道
  • Organizer: Keystone symposia on molecular and cellular biology bioactive lipids biochemistry and diseases
  • Place of Presentation: 京都市
  • Year and Date: 2010-06-07
• [Presentation] The distinct role of two arginine residues in the leukotriene A_4-specific glutathione S-transferase activity of LTC_4 synthase (2010)
  • Author(s): 齊野廣道
  • Organizer: Keystone Symposia on Molecular and Cellular Biology, Bioactive Lipids : Biochemistry and Diseases
  • Place of Presentation: 京都市
  • Year and Date: 2010-06-07
• [Presentation] プロスタグランジンE_2合成酵素1の大量発現精製と結晶化 (2009)
  • Author(s): 吾郷日出夫
  • Organizer: 日本結晶学会平成21年度年会
  • Place of Presentation: 西宮市
• [Presentation] プロスタグランジンE_2合成酵素1の大量発現精製と結晶化 (2009)
  • Author(s): 吾郷日出夫
  • Organizer: 日本結晶学会年会
  • Place of Presentation: 関西学院大学上ヶ原キャンパス(兵庫県西宮市)
• [Presentation] ロイコトリエンC_4合成酵素の変異体による酵素触媒機構の解析 (2008)
  • Author(s): 吾郷, 日出夫
  • Organizer: BMB2008
  • Place of Presentation: 神戸
  • Year and Date: 2008-12-12