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High-speed AFM observation of archaeal MCM protein complexes

Research Project

Project/Area Number 17K15433
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeMulti-year Fund
Research Field Physical pharmacy
Research InstitutionMicrobial Chemistry Research Foundation (2018)
Kanazawa University (2017)

Principal Investigator

NOSHIRO Daisuke  公益財団法人微生物化学研究会, 微生物化学研究所, 博士研究員 (90751107)

Project Period (FY) 2017-04-01 – 2019-03-31
Project Status Completed (Fiscal Year 2018)
Budget Amount *help
¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2018: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2017: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Keywords高速原子間力顕微鏡 / ヘリカーゼ / ATPase / 観察基板 / タマビジン2 / 二次元結晶 / 高速AFM / タマビジン2 / ビオチン / 生物物理 / 一分子生物学 / 生体分子 / 酵素
Outline of Final Research Achievements

Two archaeal MCM (ssoMCM and mthMCM) proteins which have helicase activities to unwind double-stranded DNA to produce single-stranded DNA were observed by high-speed AFM (HS-AFM). The observation of the full-length ssoMCM protein on mica surface revealed the formation of ladder-like long-shaped oligomers and ring (or partial ring) structures composed of more than hexamer as well as hexameric ring structure with helicase activity (or partial hexameric ring structure). In addition, observation of biotinylated full-length mthMCM protein on the 2D crystals of tamavidin 2 confirmed the formation of hexameric ring and double hexameric ring (12 mer) structures.

Academic Significance and Societal Importance of the Research Achievements

高速AFMを用いて、フレキシブルなC末端を含めた全長の古細菌由来MCMタンパク質を観察した結果、MCMタンパク質が六量体リング以外の多量体構造を形成し得ることを明らかにした。また、本研究の遂行過程で発見されたタマビジン2の二次元結晶をマイカ上に直接形成させる方法は、ビオチンを導入した生体分子の固定基板の簡便で迅速な作製法として有用であり、他の研究にも利用することができる。

Report

(3 results)
  • 2018 Annual Research Report   Final Research Report ( PDF )
  • 2017 Research-status Report
  • Research Products

    (5 results)

All 2018 2017

All Journal Article (1 results) (of which Peer Reviewed: 1 results,  Open Access: 1 results) Presentation (4 results) (of which Int'l Joint Research: 1 results)

  • [Journal Article] Substrate protein dependence of GroEL-GroES interaction cycle revealed by high-speed AFM imaging2018

    • Author(s)
      D. Noshiro and T. Ando
    • Journal Title

      Philosophical Transactions B

      Volume: 印刷中 Issue: 1749 Pages: 20170180-20170180

    • DOI

      10.1098/rstb.2017.0180

    • Related Report
      2018 Annual Research Report 2017 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] オートファジーの足場タンパク質Atg17-Atg29-Atg31複合体の高速AFM観察2018

    • Author(s)
      能代大輔、藤岡優子、野田展生、安藤敏夫
    • Organizer
      第91回日本生化学会大会
    • Related Report
      2018 Annual Research Report
  • [Presentation] 高速AFMによるAtg17-Atg29-Atg31のS字構造とAtg13-Atg17-Atg29-Atg31複合体により形成される液滴の観察2018

    • Author(s)
      能代大輔、藤岡優子、安藤敏夫、野田展生
    • Organizer
      第11回オートファジー研究会
    • Related Report
      2018 Annual Research Report
  • [Presentation] Two-dimensional crystals of tamavidin 2 for a quick and easy method of immobilization of biotinylated biomolecules2017

    • Author(s)
      能代大輔、古寺哲幸、安藤敏夫
    • Organizer
      第55回日本生物物理学会年会
    • Related Report
      2017 Research-status Report
  • [Presentation] Two-dimensional crystals of tamavidin 2 on mica surface and their application2017

    • Author(s)
      Daisuke Noshiro, Noriyuki Kodera, Toshio Ando
    • Organizer
      International Symposium on Atomic Force Microscopy at Solid-Liquid Interfaces
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research

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Published: 2017-04-28   Modified: 2020-03-30  

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