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Structural studies of a new cesium ion scavenger using a toxin having molecular switch ability

Research Project

Project/Area Number 16K14048
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Green/Environmental chemistry
Research InstitutionKyoto Institute of Technology

Principal Investigator

Kengo Kitadokoro  京都工芸繊維大学, 分子化学系, 准教授 (60283587)

Project Period (FY) 2016-04-01 – 2019-03-31
Project Status Completed (Fiscal Year 2018)
Budget Amount *help
¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
Fiscal Year 2018: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2017: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2016: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Keywords毒素 / セシウム結合タンパク質 / X線構造解析 / XSAFS / XAFS / 毒素タンパク質 / 環境材料 / セシウムイオン捕捉剤 / 分子スイッチ / マイクロカプセル
Outline of Final Research Achievements

The effective removal of the radio cesium is the problem that is urgent issues. There is not yet the development example of the remover using the property that protein couples with a cesium ion. The structure of the bacteria toxin (CPE) that an applicant analyzed structure has two metal ions binding sites in the center of the trimer. It was found that there were two glutamate clusters which have negative electric charge gathered in the molecular center of the trimer by 3D structure determination. It was revealed that these clusters have ability of various metal ions binding. It was suggested that this glutamate cluster is available to removing the cesium ion, and in this study, by using quick change methods of PCR, it was succeeded to make various mutant proteins for increasing the cesium binding property, and it was proved that some mutants can bind the cesium ion in this cluster.

Academic Significance and Societal Importance of the Research Achievements

データ測定の結果、いずれの変異体CPEにおいても約2.3 Å 分解能の回折データを収集出来た。今回、9種類の変異体帯を作成し、すべての変異体について結晶化を行い、構造決定し、構造精密化を行った。CPE-E94D(不活型含む)及びCPE-D48A-E110D(不活型のみ)については、結晶構造解析ならびにXAFS解析の両方でセシウムの結合を確認できた。XAFS解析の結果、結合している分子がセシウムであると確認できた。また、クラスター部分のアミノ酸が結合する金属イオンが、変異体の種類によって変化する可能性があることが示唆された

Report

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

    (5 results)

All 2019 2018 2016

All Journal Article (4 results) (of which Int'l Joint Research: 2 results,  Peer Reviewed: 4 results,  Open Access: 4 results) Presentation (1 results)

  • [Journal Article] Structural insights into the unique polylactate-degrading mechanism of Thermobifida alba cutinase.2019

    • Author(s)
      Kitadokoro K, Kakara M, Matsui S, Osokoshi R, Thumarat U, Kawai F, Kamitani S.
    • Journal Title

      FEBS J

      Volume: in press Issue: 11 Pages: 2087-2098

    • DOI

      10.1111/febs.14781

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Significance of a histone-like protein with its native structure for the diagnosis of asymptomatic tuberculosis2018

    • Author(s)
      Ohara Yukiko、Ozeki Yuriko、Tateishi Yoshitaka、Mashima Tsukasa、Arisaka Fumio、Tsunaka Yasuo、Fujiwara Yoshie、Nishiyama Akihito、Yoshida Yutaka、Kitadokoro Kengo、Kobayashi Haruka、Kaneko Yukihiro、Nakagawa Ichiro、Maekura Ryoji、Yamamoto Saburo、Katahira Masato、Matsumoto Sohkichi
    • Journal Title

      PLOS ONE

      Volume: 13 Issue: 10 Pages: 0204160-0204160

    • DOI

      10.1371/journal.pone.0204160

    • NAID

      120006534592

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Staphylococcus aureus lipase: purification, kinetic characterization, crystallization and crystallographic study2018

    • Author(s)
      Tanaka M, Kamitani S, Kitadokoro K.
    • Journal Title

      Acta Crystallographica Section F

      Volume: 74 Issue: 9 Pages: 567-570

    • DOI

      10.1107/s2053230x18010506

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Expression, Purification and Crystallization of Thermostable Mutant of Cutinase Est1 from Thermobifida alba2018

    • Author(s)
      Kitadokoro K, Matsui S, Oshkosh R, Nakata K, Thumarat U, Kawai F, Kamitani S.
    • Journal Title

      Advances in Bioscience and Biotechnology

      Volume: 9 Issue: 05 Pages: 215-223

    • DOI

      10.4236/abb.2018.95015

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Presentation] 分子スイッチ能を有する毒素を利用した新規なセシウムイオン捕捉剤の作製2016

    • Author(s)
      高田大暉、堀口安彦、北所健悟
    • Organizer
      第6回4大学連携研究フォーラム
    • Place of Presentation
      京都薬科大学
    • Related Report
      2016 Research-status Report

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Published: 2016-04-21   Modified: 2020-03-30  

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