Elucidation of catalytic mechanism of two characteristic enzymes and its application based on crystal structures.

2014 Fiscal Year Final Research Report

• Project/Area Number: 24570130
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Structural biochemistry
• Research Institution: Kyoto University
• Principal Investigator: FUJIHASHI Masahiro 京都大学, 理学(系)研究科(研究院), 助教 (10397581)
• Co-Investigator(Renkei-kenkyūsha): SATO Tsutomu 新潟大学, 農学部, 准教授 (80334655) ; ISHIDA Toyokazu 独立行政法人産業技術総合研究所, ナノシステム研究部門, 研究員 (70443166)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Keywords: 基質の歪み / 脱炭酸反応 / 結晶構造解析 / 高分解能結晶構造 / 計算機シミュレーション / 酵素反応機構

Outline of Final Research Achievements

This project targeted two enzymes. Regarding the first enzyme ODCase, we determined about 10 crystal structures using various mutants and/or ligands. The best resolution of the determined structures is 1.03 A, which is much better than those of the over 200 determined ODCase structures thus far. Based on the structures with biochemical and computational analyses, we elucidated that substrate distortion contributes to 10-15% free energy decrease of the transition state by ODCase. Regarding the other enzyme Cyc2, we elucidated the enzyme is not appropriate for the crystallographic analysis. We are now attempt to purify and crystallize a homologous protein from a different organism.

Free Research Field

構造生物学