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Development of reverse-protein splicing method using proteases

Research Project

Project/Area Number 18K05113
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Review Section Basic Section 33020:Synthetic organic chemistry-related
Research InstitutionTokai University

Principal Investigator

Katayama Hidekazu  東海大学, 工学部, 准教授 (30580857)

Project Period (FY) 2018-04-01 – 2021-03-31
Project Status Completed (Fiscal Year 2020)
Budget Amount *help
¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2020: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2019: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2018: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
Keywordsタンパク質逆スプライシング / プロテアーゼ / ネイティブケミカルライゲーション / タンパク質逆スプライシング反応 / ペプチド縮合反応 / Native chemical ligation / ペプチドライゲーション / プロテーゼ
Outline of Final Research Achievements

Protein splicing is found in nature, although its reverse reaction has not yet been developed. In this project, we tried to develop reverse-protein splicing (RPS) reaction using a protease as catalyst. When using lysyl endopeptidase (LEP) as the catalyst, RPS reaction proceeded in ethylene glycol aqueous solution, but the yield of desired product was low. The peptide with non-proteinogenic amino acid was also acted as an insert in this reaction. This is the first case for one-pot chemical RPS reaction, and it might be useful for the regioselective chemical modification of polypeptides.

Academic Significance and Societal Importance of the Research Achievements

有機化学的にRPS反応を行った例は、これまでに報告されていないことから、世界初の実施例であると考えられる。この方法を利用すれば、将来的に組換えタンパク質の部位特異的なアミノ酸配列変換などに利用できるものと期待される。また、非タンパク質性の化学構造も挿入可能であったことから、蛍光団の挿入などの部位特異的な化学修飾にも応用可能であり、新たなタンパク質化学修飾法として生化学分野、ケミカルバイオロジー分野への応用が期待される。

Report

(4 results)
  • 2020 Annual Research Report   Final Research Report ( PDF )
  • 2019 Research-status Report
  • 2018 Research-status Report
  • Research Products

    (9 results)

All 2020 2019 2018

All Journal Article (4 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 4 results,  Open Access: 1 results) Presentation (5 results) (of which Int'l Joint Research: 1 results)

  • [Journal Article] A relaxin-like gonad-stimulating peptide identified from the starfish Astropecten scoparius2020

    • Author(s)
      Mita Masatoshi、Osugi Tomohiro、Matsubara Shin、Kawada Tsuyoshi、Satake Honoo、Katayama Hidekazu
    • Journal Title

      Molecular Reproduction and Development

      Volume: 88 Issue: 1 Pages: 34

    • DOI

      10.1002/mrd.23444

    • URL

      https://localhost/en/publications/f02472a4-e280-4797-8af1-8a183bb59c60

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Application of 2,2′‐dipyridyl disulfide‐mediated thiazolidine ring‐opening reaction to glycoprotein synthesis: Total chemical synthesis of evasin‐32020

    • Author(s)
      Katayama Hidekazu、Nagata Koji
    • Journal Title

      Journal of Peptide Science

      Volume: 27 Issue: 2

    • DOI

      10.1002/psc.3290

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed
  • [Journal Article] A novel approach for preparing disulfide-rich peptide-KLH conjugate applicable to the antibody production2019

    • Author(s)
      Katayama Hidekazu、Mizuno Ryo、Mita Masatoshi
    • Journal Title

      Bioscience, Biotechnology, and Biochemistry

      Volume: 83 Issue: 10 Pages: 1791

    • DOI

      10.1080/09168451.2019.1618696

    • URL

      https://pure.teikyo.jp/en/publications/e887db41-d536-4ae8-953a-2833a32a1b98

    • Related Report
      2019 Research-status Report
    • Peer Reviewed
  • [Journal Article] Chemical synthesis of N-glycosylated insulin-like androgenic gland factor from the freshwater prawn Macrobrachium rosenbergii2019

    • Author(s)
      Katayama Hidekazu、Nagasawa Hiromichi
    • Journal Title

      Journal of Peptide Science

      Volume: 25 Issue: 11

    • DOI

      10.1002/psc.3215

    • URL

      https://pure.teikyo.jp/en/publications/01e56830-9c51-4fcf-a247-a94f83b5510e

    • Related Report
      2019 Research-status Report
    • Peer Reviewed
  • [Presentation] Chemical synthesis of insulin-like androgenic gland factor from the crayfish Procambarus clarkii2020

    • Author(s)
      片山 秀和、豊田 賢治、田中 陽菜、大平 剛
    • Organizer
      第57回ペプチド討論会
    • Related Report
      2020 Annual Research Report
  • [Presentation] 新規のチアゾリジン開環反応を利用した糖タンパク質Evasin-3の化学合成2020

    • Author(s)
      片山秀和、永田宏次
    • Organizer
      日本農芸化学会2020年度大会
    • Related Report
      2019 Research-status Report
  • [Presentation] Chemical synthesis of insulin-like androgenic gland factor from the freshwater prawn Macrobrachium rosenbergii2019

    • Author(s)
      片山秀和、長澤寛道
    • Organizer
      第56回ペプチド討論会
    • Related Report
      2019 Research-status Report
  • [Presentation] Application of the novel thiazolidine ring opening reaction to glycoprotein synthesis2018

    • Author(s)
      Hidekazu Katayama
    • Organizer
      10th International Peptide Symposium
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research
  • [Presentation] マヒトデ由来リラキシン様ペプチドに対する抗体作製に向けた誘導体の合成研究2018

    • Author(s)
      片山秀和、水野涼、三田雅敏
    • Organizer
      第43回日本比較内分泌学会
    • Related Report
      2018 Research-status Report

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Published: 2018-04-23   Modified: 2022-01-27  

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