Physicochemical study and drug development for an unknown compound binding to metallo-beta-lactamase

• Project/Area Number: 24790045
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Physical pharmacy
• Research Institution: Kumamoto University
• Principal Investigator: YOSHIHIRO Yamaguchi 熊本大学, 環境安全センター, 准教授 (10363524)
• Project Period (FY): 2012-04-01 – 2014-03-31
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2013: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2012: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
• Keywords: 蛋白質の発色 / 薬剤耐性菌 / 酵素 / 酵素阻害剤 / β-ラクタム剤 / Zn酵素 / 未知化合物 / タンパク質から分離 / 国際情報交換 / 阻害剤 / X線結晶構造解析 / 速度論的解析

Research Abstract

Metallo-beta-lactamase (MBL) is an enzyme capable of hydrolyzing nearly all beta-lactam antibiotics. MBLs are normally colorless, however a series of red-violet MBL enzymes were discovered. The cause of this coloration was presumed to be due to colored compounds bound to the MBL. This research is an attempt to isolate these colored MBLs and examine their structure and binding mechanisms.
It was assumed that denaturing the MBL enzyme would cause separation from the colored compound. This study confirmed that these colored compounds degrade easily with denaturation of the MBL enzyme.
Future research will involve finding the molecular weight of the compound via mass spectrometry and other analyses to further understand its chemical structure and binding mechanism.

Research Products

• [Journal Article] Crystallization and Preliminary X-ray Analysis of the Subclass B3 Metallo-β-Lactamase SMB-1 that Confers Carbapenem Resistance (2012)
  • Author(s): J.Wachino, et al
  • Journal Title: Acta Cryst Volume: F68 Issue: 3 Pages: 343-346
  • DOI: 10.1107/s1744309112004691
  • Peer Reviewed
• [Journal Article] Structural Insights into the Subclass B3 Metallo-β-Lactamase SMB-1 and the Mode of Inhibition by the Common Metallo-β-Lactamase Inhibitor Mercaptoacetate (2012)
  • Author(s): Jun-ichi Wachino, Yoshihiro Yamaguchi, Shigetarou Mori, Hiromasa Kurosaki, Yoshichika Arakawa, Keigo Shibayama
  • Journal Title: Antimicrob. Agents Chemother. Volume: 57 Issue: 1 Pages: 101-109
  • DOI: 10.1128/aac.01264-12
  • Peer Reviewed
• [Presentation] 金属酵素メタロ-β-ラクタマーゼに結合した未知化合物の分離法の探索 (2015)
  • Author(s): 山口佳宏、甲斐紀子、稲津 誠、荒川宜親、黒崎博雅
  • Organizer: 日本蛋白質科学会
  • Place of Presentation: 横浜
• [Presentation] 金属酵素メタロ-β-ラクタマーゼに結合した未知化合物の分離法の探索 (2014)
  • Author(s): 山口佳宏、甲斐紀子、稲津誠、荒川宜親、黒崎博雅
  • Organizer: 2014年度日本蛋白質科学会
  • Place of Presentation: 横浜産貿ホールマリネリア
• [Presentation] e-Learning in Environmental and Safety Education at Kumamoto University (2012)
  • Author(s): Yoshihiro Yamaguchi
  • Organizer: SETAC Asia Pacific 2012
  • Place of Presentation: ANA Hotel Kumamoto New Sky, Kumamoto, Japan
• [Presentation] Citrobacter freundii由来メタロ-β-ラクタマーゼ（KHM-1）の単核Zn型および複核Zn型の結晶構造 (2012)
  • Author(s): 山口佳宏、西並 隆、安田健二、切替照雄、山縣ゆり子、黒崎博雅
  • Organizer: 第12回日本蛋白質科学会
  • Place of Presentation: 名古屋国際会議場、名古屋
• [Presentation] 効率よくセフェム系βラクタム剤を加水分解するメタロ-β-ラクタマーゼ（KHM-1）の構造学的解 (2012)
  • Author(s): 山口佳宏、西並 隆、安田健二、切替照雄、山縣ゆり子、黒崎博雅
  • Organizer: 第10回次世代を担う若手のためのフィジカル・ファーマ・フォーラム
  • Place of Presentation: ホテル平安の森京都、京都
• [Remarks]
  • URL: http://yamaguchi-labo.jp
• [Remarks] 山口ラボ
  • URL: http://yamaguchi-labo.jp
• [Remarks] 酵素機能化学 山口研究室
  • URL: http://www.yamaguchi-labo.jp/
• [Remarks] 熊本大学環境安全センター
  • URL: http://www.esc.kumamoto-u.ac.jp/