• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

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

Research Project

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
Project Status Completed (Fiscal Year 2019)
Budget Amount *help
¥17,680,000 (Direct Cost: ¥13,600,000、Indirect Cost: ¥4,080,000)
Fiscal Year 2018: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2017: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2016: ¥9,360,000 (Direct Cost: ¥7,200,000、Indirect Cost: ¥2,160,000)
Keywords酵素 / X線結晶構造解析 / 結晶構造 / プロトン化状態 / 酵素反応 / 構造生物学 / タンパク質 / 結晶構造解析 / 基質の歪み / オロチジン一リン酸脱炭酸酵素
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.

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。

Report

(4 results)
  • 2019 Final Research Report ( PDF )
  • 2018 Annual Research Report
  • 2017 Annual Research Report
  • 2016 Annual Research Report
  • Research Products

    (14 results)

All 2018 2017 2016 Other

All Int'l Joint Research (2 results) Presentation (12 results) (of which Int'l Joint Research: 3 results,  Invited: 5 results)

  • [Int'l Joint Research] Toronto General Research Institute/Ontario Cancer Institute/University of Toronto(カナダ)

    • Related Report
      2017 Annual Research Report
  • [Int'l Joint Research] Ontario Cancer Institute/Toronto General Research Institute(Canada)

    • Related Report
      2016 Annual Research Report
  • [Presentation] Determination and application of enzyme structure2018

    • Author(s)
      Masahiro Fujihashi
    • Organizer
      Scientific Symposium in Honor of Professor Emil F. Pai: Why Structure Matters
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] 生合成酵素の立体構造とその利用2018

    • Author(s)
      藤橋雅宏
    • Organizer
      新潟大学 ケミカルバイオロジー共用ユニット主催第3回技術セミナー
    • Related Report
      2018 Annual Research Report
    • Invited
  • [Presentation] 酵素の結晶構造解析とその利用2018

    • Author(s)
      藤橋雅宏
    • Organizer
      東大薬学部特別講演会
    • Related Report
      2017 Annual Research Report
    • Invited
  • [Presentation] 生合成酵素の立体構造とその利用2018

    • Author(s)
      藤橋雅宏
    • Organizer
      新潟大学 第三回技術セミナー
    • Related Report
      2017 Annual Research Report
    • Invited
  • [Presentation] Determination and application of enzyme structure2018

    • Author(s)
      Masahiro Fujihashi
    • Organizer
      Scientific Symposium in Honor of professor emil F. Pai
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 全自動測定ビームタイムを使ってみて2018

    • Author(s)
      藤橋雅宏
    • Organizer
      第3回 タンパク質結晶構造解析ビームライン中級者向け講習会
    • Related Report
      2017 Annual Research Report
    • Invited
  • [Presentation] オロチジン一リン酸脱炭酸酵素の活性中心における負電荷反発の解明2018

    • Author(s)
      岩原卓哉, Emil F. Pai, Lakshmi P. Kotra, 三木邦夫, 藤橋雅宏
    • Organizer
      日本結晶学会 2018年度年会
    • Related Report
      2017 Annual Research Report
  • [Presentation] オロチジン一リン酸脱炭酸酵素の酵素基質複合体における基質歪みの由来の分析2017

    • Author(s)
      藤橋雅宏, Emil F. Pai, Lakshmi P. Kotra, 三木邦夫
    • Organizer
      第17回日本蛋白質科学会年会
    • Related Report
      2017 Annual Research Report
  • [Presentation] 取得効率やX線損傷を考慮したデータ収集例2017

    • Author(s)
      藤橋雅宏
    • Organizer
      第2回 タンパク質結晶構造解析ビームライン中級者向け講習会
    • Related Report
      2017 Annual Research Report
  • [Presentation] オロチジン一リン酸脱炭酸酵素の活性中心における静電反発の解析2017

    • Author(s)
      岩原卓哉, 藤橋雅宏, Emil F. Pai, Lakshmi P. Kotra, 三木邦夫
    • Organizer
      日本結晶学会 2017年度年会
    • Related Report
      2017 Annual Research Report
  • [Presentation] オロチジン一リン酸脱炭酸酵素の反応時における基質歪みの由来2016

    • Author(s)
      藤橋雅宏,石田豊和,Emil F. Pai,Lakshmi P. Kotra,三木邦夫
    • Organizer
      日本結晶学会平成28年度年会
    • Place of Presentation
      茨城県立県民文化センター, 茨城県水戸市
    • Year and Date
      2016-11-17
    • Related Report
      2016 Annual Research Report
  • [Presentation] Contribution of the substrate distortion to the reaction of orotidine 5’-monophosphate decarboxylase2016

    • Author(s)
      Masahiro Fujihashi, Toyokazu Ishida, Lakshmi P. Kotra, Emil F. Pai, and Kunio Miki
    • Organizer
      The Fifth International Conferencee on Cofactors & Active Enzyme Molecule 2016
    • Place of Presentation
      Unazuki, Toyama, Japan
    • Related Report
      2016 Annual Research Report
    • Int'l Joint Research

URL: 

Published: 2016-04-21   Modified: 2021-02-19  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi