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Development of a method to grow large protein crystals for neutron structural analysis

Research Project

Project/Area Number 16K14677
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Structural biochemistry
Research InstitutionHokkaido University

Principal Investigator

Yao Min  北海道大学, 先端生命科学研究院, 教授 (40311518)

Co-Investigator(Renkei-kenkyūsha) Kato Koji  北海道大学, 先端生命科学研究院, 助教 (30452428)
Research Collaborator ADACHI Motoyasu  国立研究開発法人日本原子力研究開発機構, 原子力科学研究部門・量子ビーム応用研究センター, グループリーダー (60293958)
Project Period (FY) 2016-04-01 – 2018-03-31
Project Status Completed (Fiscal Year 2017)
Budget Amount *help
¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
Fiscal Year 2017: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Keywords中性子線結晶構造解析 / タンパク質結晶成長 / LCST型温度応答ポリマー / ゲル / ゾル / X線結晶構造解析 / 結晶成長エントロピー / 相転移 / 水分子 / 温度応答ポリマー / 中性子線結晶解析
Outline of Final Research Achievements

The purpose of this project is to establish technology for growing large protein crystals with high quality, which is essential to analyze the protein structure by neutron crystallography. We tried to grow up lager crystals by using LCST-type polymers for several protein samples, and have succeeded in growing lager crystals. Among those samples, we collected neutron diffraction data of GatCAB (Mw:110 kDa), and determined the neutron structure. The effectiveness of LCST-type polymer on the growth of large protein crystal was confirmed. Moreover, the entropy changes during in the crystallization solution using LCST-type polymer were investigated during crystal growth. Therefore, it is assumed that LCST-type polymer changed the state of water molecules around the protein molecules.

Report

(3 results)
  • 2017 Annual Research Report   Final Research Report ( PDF )
  • 2016 Research-status Report
  • Research Products

    (5 results)

All 2017 2016 Other

All Presentation (4 results) (of which Int'l Joint Research: 1 results) Remarks (1 results)

  • [Presentation] Challenging to visualize ammonia transposition in a channel of amidotransferase GatCAB using neutron macromolecular crystallography2017

    • Author(s)
      Min Yao, Long Li
    • Organizer
      第55回日本生物物理学会年会
    • Related Report
      2017 Annual Research Report
  • [Presentation] Attempt to elucidate ammonia transportation of GatCAB by NMC2017

    • Author(s)
      Long Li, Koji Kato, Akira Shinoda, Toyoyuki Ose, Min Yao
    • Organizer
      平成29年度日本結晶学会年会
    • Related Report
      2017 Annual Research Report
  • [Presentation] Heterogeneous Nucleation of Protein Crystals by using Nucleants2016

    • Author(s)
      Long Li, Akira Shinoda, Koji Kato, Min Yao
    • Organizer
      The 4th International Life-Science Symposium (ILSS)
    • Place of Presentation
      Hokkaido University Conference Hall(北海道・札幌市)
    • Year and Date
      2016-11-18
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research
  • [Presentation] Epitaxial Nucleation of Protein Crystals on the Crystalline Surface2016

    • Author(s)
      李龍,篠田晃,加藤公児,姚閔
    • Organizer
      平成28年度日本結晶学会年会
    • 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 Research-status Report
  • [Remarks] 研究室のホームページ

    • URL

      http://altair.sci.hokudai.ac.jp/g6/

    • Related Report
      2017 Annual Research Report

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Published: 2016-04-21   Modified: 2019-03-29  

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