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Crystallography of membrane protein complex by S-SAD method

Research Project

Project/Area Number 15K14468
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Structural biochemistry
Research InstitutionHigh Energy Accelerator Research Organization

Principal Investigator

KATO Ryuichi  大学共同利用機関法人高エネルギー加速器研究機構, 物質構造科学研究所, 准教授 (50240833)

Co-Investigator(Renkei-kenkyūsha) MAKIYO Hisayoshi  高エネルギー加速器研究機構, 物質構造科学研究所, 研究員 (00467474)
MATSUGAKI Naohiro  高エネルギー加速器研究機構, 物質構造科学研究所, 准教授 (50342598)
YAMADA Yusuke  高エネルギー加速器研究機構, 物質構造科学研究所, 助教 (20391708)
MANYA Hiroshi  東京都健康長寿医療センター研究所, 研究副部長 (20321870)
Project Period (FY) 2015-04-01 – 2018-03-31
Project Status Completed (Fiscal Year 2017)
Budget Amount *help
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2017: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2016: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2015: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Keywords筋ジストロフィー / ジストログリカン / 膜タンパク質 / 結晶構造解析 / POMT1 / POMT2
Outline of Final Research Achievements

For structural study of the membrane protein complex POMT1-POMT2, which is involved in congenital muscular dystrophy, expression experiments in mammalian cultured cells were performed. Unfortunately, the expression level was very low, which is insufficient to start crystallization experiments. We also examined the method of crystallizing proteins expressed in insect body, but the proteins did not express.
In parallel, we studied development of a method of performing X-ray crystal structure analysis of membrane protein POMT1-POMT2 by a phase determination method utilizing the anomalous dispersion effect of sulfur atoms (S-SAD method). By using standard proteins with synchrotron beamline at KEK, S-SAD method was improved, and sufficient results were obtained.

Report

(4 results)
  • 2017 Annual Research Report   Final Research Report ( PDF )
  • 2016 Research-status Report
  • 2015 Research-status Report
  • Research Products

    (6 results)

All 2017 2016

All Journal Article (2 results) (of which Peer Reviewed: 2 results,  Open Access: 2 results) Presentation (4 results)

  • [Journal Article] Experimental phase determination with selenomethionine or mercury-derivatization in serial femtosecond crystallography.2017

    • Author(s)
      Yamashita, K., et al.
    • Journal Title

      IUCrJ

      Volume: 4 Issue: 5 Pages: 639-647

    • DOI

      10.1107/s2052252517008557

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] In-situ data collection at the photon factory macromolecular crystallography beamlines.2016

    • Author(s)
      Yamada, Y., Hiraki, M., Matsugaki, N., Kato, R., Senda, T.
    • Journal Title

      AIP Conf. Proc.

      Volume: 1741 Pages: 050023-050023

    • DOI

      10.1063/1.4952943

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] ヒトタンパク質の哺乳動物を用いた細胞内結晶化2017

    • Author(s)
      小祝孝太郎、月本準、東哲也、真板宣夫、山田悠介、平木雅彦、加藤龍一、Leonard M. G. Chavas、千田俊哉、伊藤孝司、湯本史明
    • Organizer
      第17回日本蛋白質科学会年会
    • Related Report
      2017 Annual Research Report
  • [Presentation] BINDSにおけるPFタンパク質結晶構造解析ビームラインの支援と高度化2017

    • Author(s)
      山田悠介、引田理英、松垣直宏、平木雅彦、田辺幹雄、湯本史明、加藤龍一、千田俊哉
    • Organizer
      日本結晶学会平成29年度年会
    • Related Report
      2017 Annual Research Report
  • [Presentation] PFタンパク質結晶構造解析ビームラインによる構造生物学研究支援2017

    • Author(s)
      山田 悠介、松垣 直宏、引田 理英、平木 雅彦、加藤 龍一、千田 俊哉
    • Organizer
      ConBio2017
    • Related Report
      2017 Annual Research Report
  • [Presentation] ヒトタンパク質の哺乳動物細胞を用いた細胞内結晶化2017

    • Author(s)
      小祝 孝太郎、月本 準、東 哲也、真板 宣夫、山田 悠介、平木 雅彦、加藤 龍一、千田 俊哉、Chavas Leonard、伊藤 孝司、湯本 史明
    • Organizer
      ConBio2017
    • Related Report
      2017 Annual Research Report

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Published: 2015-04-16   Modified: 2019-03-29  

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