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2019 Fiscal Year Final Research Report

Experimental analysis of the electrostatic repulsion between enzyme and substrates to elucidate the reaction mechanism, and development anti-malaria agent

Research Project

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Project/Area Number 16H04751
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Structural biochemistry
Research InstitutionKyoto University

Principal Investigator

Fujihashi Masahiro  京都大学, 理学研究科, 助教 (10397581)

Project Period (FY) 2016-04-01 – 2019-03-31
Keywords酵素 / X線結晶構造解析
Outline of Final Research Achievements

In order to elucidate the details of the destabilization of the enzyme-substrate complex, we established a base for high-resolution X-ray crystallographic analysis while avoiding the influence of crystal damage by X-rays. First, we enabled large-scale sample preparation comparing to several times larger than conventional ones. We then identified the crystal form which is the most suitable for analysis and established the protocol for its preparation. Also, we analyzed the relationship between the absorbed X-ray dose of the crystal and the damage of the internal structure of the enzyme. The analysis enabled us to collect damage-less diffraction dataset. The best resolution of the obtained dataset was 0.99 angstrom.

Free Research Field

構造生物学

Academic Significance and Societal Importance of the Research Achievements

酵素基質複合体の不安定化は、理論的には全ての酵素が利用可能な機構であるにもかかわらず、実験的な解析例はほとんど無い。また、酵素と基質の間には引力が働くと考えるのが常識であり、静電的な反発が活性に関わるとの報告は存在しない。本研究はこのように、理論的に可能でありながら、これまでに報告例のない機構の解明を目指して、これに必要な試料の調製方法の確立、データ取得手法の検討、ならびにデータ取得などを行った。

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Published: 2021-02-19  

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