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Analysis of abnormal prion protein core structure by proteolysis at high temperature

Research Project

Project/Area Number 15K07392
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Applied biochemistry
Research InstitutionOsaka University

Principal Investigator

Koga Yuichi  大阪大学, 工学研究科, 准教授 (30379119)

Project Period (FY) 2015-04-01 – 2018-03-31
Project Status Completed (Fiscal Year 2017)
Budget Amount *help
¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2017: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2016: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2015: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Keywordsプロテアーゼ / 蛋白質工学 / PrP / Subtilisin / 耐熱性酵素 / 超好熱菌 / プリオン / プリオンタンパク質
Outline of Final Research Achievements

A hyperthermostable protease, Tk-subtilisin, can degrade PrPSc. Scrapie-infected mouse brain homogenate ( RML strain ) was degraded by Tk-subtilisin under high-temperature condition and the degradation products were inoculated into culture cell. At high-temperature condition, Tk-subtilisin degraded PrP from RML homogenate, whereas proteinase K ( PK ) hardly degraded. In addition, Western blotting of the cell lysate inoculated the degradation product showed that the amount of PrPSc decreased significantly. Moreover, the signals of PrPSc were not observed when the concentration of Tk-subtilisin was increased. These results suggest that the combination of heat treatment and proteolysis by Tk-subtilisin is effective in reducing the infectivity titer of PrPSc.

Report

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

    (14 results)

All 2017 2016 2015

All Journal Article (1 results) (of which Peer Reviewed: 1 results,  Open Access: 1 results,  Acknowledgement Compliant: 1 results) Presentation (12 results) (of which Int'l Joint Research: 6 results,  Invited: 4 results) Book (1 results)

  • [Journal Article] Effects of surfactant and a hyperthermostable protease on infectivity of Scrapie-infected mouse brain homogenate.2015

    • Author(s)
      Hirata, A., Sakudo, A., Takano, K., Kanaya, S. and Koga, Y.
    • Journal Title

      Journal of Biotechnol and Biomater

      Volume: 5 Issue: 03 Pages: 194-194

    • DOI

      10.4172/2155-952x.1000194

    • Related Report
      2015 Research-status Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Presentation] 超好熱菌由来Thermococcus kodakarensis KOD1 由来subtilisin 様プロテアーゼとその利用2017

    • Author(s)
      古賀雄一、清水七海、足立航平、大政健史
    • Organizer
      日本農芸化学会
    • Place of Presentation
      京都女子大学
    • Year and Date
      2017-03-17
    • Related Report
      2016 Research-status Report
  • [Presentation] βサンドイッチドメインを用いた結合分子骨格の検討2017

    • Author(s)
      来住秀憲、杉田慎之介、大政健史、古賀雄一
    • Organizer
      第17回日本蛋白質科学会
    • Related Report
      2017 Annual Research Report
  • [Presentation] 超好熱古細菌由来プロテアーゼTk0076の成熟化機構解明を目指したプロペプチドの探索2017

    • Author(s)
      田口萌恵、大政健史、古賀雄一
    • Organizer
      第17回日本蛋白質科学会
    • Related Report
      2017 Annual Research Report
  • [Presentation] Construction of large-scale expression system by suppressing the maturation of Tk-SP from hyperthermophilic archaeon2017

    • Author(s)
      前川大樹、古賀雄一、大政 健史
    • Organizer
      第49回化学工学会秋季大会
    • Related Report
      2017 Annual Research Report
  • [Presentation] Subtilisin-like proteases from a hyper-thermophilic archaeon and their application2017

    • Author(s)
      古賀雄一
    • Organizer
      BIOTEC-OSAKA Univ. Special Seminar
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] 超好熱菌由来酵素の構造機能相関研究とその応用2017

    • Author(s)
      古賀雄一
    • Organizer
      第69回日本生物工学会大会
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Protein engineering on a Subtilisin-like protease from a hyper-thermophilic archaeon2017

    • Author(s)
      古賀雄一
    • Organizer
      The 29th Annual Meeting of the Thai Society for Biotechnology and International Conference
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] 生物資源を活用した研究事例2017

    • Author(s)
      古賀雄一
    • Organizer
      生物資源活用の世界展開~グローカルバイオテクノロジー~シンポジウム
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] ケラチン分解酵素の機能不明C末端ドメインの機能解析2016

    • Author(s)
      杉田 愼之助、 大政 健史、 古賀 雄一
    • Organizer
      第68回日本生物工学会大会
    • Place of Presentation
      富山国際会議場
    • Year and Date
      2016-09-28
    • Related Report
      2016 Research-status Report
  • [Presentation] Hyper-Thermophilic Subtilisin-like Proteases from Thermococcus kodakarensis and Their Application2016

    • Author(s)
      Yuichi Koga, Ryo Uehara, Kazufumi Takano
    • Organizer
      Extremophiles 2016
    • Place of Presentation
      京都大学
    • Year and Date
      2016-09-12
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research
  • [Presentation] Construction of a Novel Expression System of Toxic Protease from a Hyperthermophilic Archaeon Thermococcus kodakararensis KOD12016

    • Author(s)
      Daiki Maegawa1, Kohei Adachi1, Takeshi Omasa1, Yuichi Koga
    • Organizer
      Extremophiles 2016
    • Place of Presentation
      京都大学
    • Year and Date
      2016-09-12
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research
  • [Presentation] Intein を利用した超好熱菌由来プロテアーゼ(Tk-SP)の発現系構築2016

    • Author(s)
      足立航平、上原了、Tannous E. Antonios、大政健史、古賀雄一
    • Organizer
      日本農芸化学会
    • Place of Presentation
      札幌コンベンションセンター(北海道・札幌)
    • Year and Date
      2016-03-27
    • Related Report
      2015 Research-status Report
  • [Book] Biotechnology of Microbial Enzymes, production, biocatalysis and industrial applications2016

    • Author(s)
      Uehara, R., Takano, K., Kanaya, S. and Koga, Y.
    • Total Pages
      608
    • Publisher
      Academic Press
    • Related Report
      2016 Research-status Report

URL: 

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

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