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Structural analysis of active site in a photoprotein (pholasin) and its application to bright imaging

Research Project

Project/Area Number 19K05844
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Review Section Basic Section 38040:Bioorganic chemistry-related
Research InstitutionKobe University

Principal Investigator

KUSE MASAKI  神戸大学, 農学研究科, 教授 (40335013)

Project Period (FY) 2019-04-01 – 2022-03-31
Project Status Completed (Fiscal Year 2021)
Budget Amount *help
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2021: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2020: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2019: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Keywords生体成分 / 生理活性 / 蛋白質 / 有機化学 / 生物発光 / タンパク質 / 生体分子
Outline of Research at the Start

以下の年度計画で順次計画を進める。
平成31年度:①13C標識DCLの合成と再構成フォラシンのNMR測定、②N/O置換DCL誘導体の合成
平成32年度:①フォラシンの再構成と構造活性相関、②NMR、ラマン・赤外分光によるフォラシンの解析
平成33年度:①高輝度フォラシンの創製、②発現アポフォラシンから再構成したフォラシンのX線結晶構造解析

Outline of Final Research Achievements

This research of a luminescent protein (pholasin) began with preparing apoproteins without substrates. Then, using the gene-expressed apopholasin and substrate, we found the conditions for activating the reconstituted pholasin. Using these conditions, we found a substrate structure that emits light brighter than the natural substrate. Subsequently, we were able to collect information on the structure and luminescent activity of the substrate through structure-activity relationship studies. This result enabled us to create artificial pholasin, which emitted highly luminescent light and established a fundamental technology for visualizing reactive oxygen species (ROS). Furthermore, we found a method for introducing a linker into the substrate to modify the emission wavelength. These results have enabled applications using various wavelengths.

Academic Significance and Societal Importance of the Research Achievements

活性酸素種(ROS)は細胞内におけるシグナル伝達において、セカンドメッセンジャーとして重要な役割を果たしている。様々なROS検出法が確立されているが、ROSを時間・空間で可視化する手段がないため、未だに不明な点も多いのが現状である。本研究で確立した発光タンパク質フォラシンを用いた高輝度発光系は、ROS検出のための手段として利用することができることから、ROSに関連するシグナル伝達の研究分野において利用されることが期待される。さらに発光波長を改変させる可能性もあることから、生体組織中でのROSの可視化手段としての利用も期待される意義がある。

Report

(4 results)
  • 2021 Annual Research Report   Final Research Report ( PDF )
  • 2020 Research-status Report
  • 2019 Research-status Report
  • Research Products

    (15 results)

All 2022 2021 2020 2019 Other

All Int'l Joint Research (2 results) Journal Article (8 results) (of which Int'l Joint Research: 4 results,  Peer Reviewed: 8 results,  Open Access: 1 results) Presentation (4 results) (of which Invited: 1 results) Remarks (1 results)

  • [Int'l Joint Research] Knight Scientific Ltd/Nottingham University(英国)

    • Related Report
      2021 Annual Research Report
  • [Int'l Joint Research] Knight Scientific Ltd/Nottingham University(英国)

    • Related Report
      2020 Research-status Report
  • [Journal Article] Phenylpropanoids and neolignans isolated from Myristica fragrans enhance glucose uptake in myotubes2022

    • Author(s)
      Yoshioka Yasukiyo、Kono Ryunoshin、Kuse Masaki、Yamashita Yoko、Ashida Hitoshi
    • Journal Title

      Food & Function

      Volume: 13 Issue: 7 Pages: 3879-3893

    • DOI

      10.1039/d1fo04408g

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Isolation and identification of metabolites from ethyl acetate leaf extract of Solenostemma argel2021

    • Author(s)
      Hanaa Hassabelrasoul, Maiko Moriguchi, Bubwoong Kang, Awatif Ahmed Siribel, Masaki Kuse*
    • Journal Title

      Agriculture and Natural Resources

      Volume: 55 Issue: 5 Pages: 757-763

    • DOI

      10.34044/j.anres.2021.55.5.06

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Anti-allergic property of 4,8-sphingadienine stereoisomers in vivo and in vitro model2021

    • Author(s)
      Shimizu Haruka、Kuse Masaki、Minato Ken-ichiro、Mizuno Masashi
    • Journal Title

      Biochemical and Biophysical Research Communications

      Volume: 577 Pages: 32-37

    • DOI

      10.1016/j.bbrc.2021.08.071

    • NAID

      120007148940

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Expression of recombinant apopholasin using a baculovirus-silkworm multigene expression system and activation via dehydrocoelenterazine2020

    • Author(s)
      Maiko Moriguchi, RyoTakahashi, Bubwoong Kang, Masaki Kuse
    • Journal Title

      Bioorganic & Medicinal Chemistry Letters

      Volume: 30 Issue: 12 Pages: 127177-127177

    • DOI

      10.1016/j.bmcl.2020.127177

    • NAID

      120006864548

    • Related Report
      2020 Research-status Report 2019 Research-status Report
    • Peer Reviewed
  • [Journal Article] Marine Bioluminescence with Dehydrocoelenterazine, an Imidazopyrazinone Compound.2020

    • Author(s)
      Masaki Kuse
    • Journal Title

      Topics in Heterocyclic Chemistry

      Volume: 58 Pages: 1-19

    • DOI

      10.1007/7081_2020_41

    • ISBN
      9789811646362, 9789811646379
    • Related Report
      2020 Research-status Report
    • Peer Reviewed
  • [Journal Article] Concise synthesis of heliolactone, a non-canonical strigolactone isolated from sunflower2020

    • Author(s)
      Shunya Yamamoto, Taiki Atarashi, Masaki Kuse, Yukihiro Sugimoto, Hirosato Takikawa
    • Journal Title

      Biosci. Biotechnol. Biochem

      Volume: 84 Issue: 6 Pages: 1113-1118

    • DOI

      10.1080/09168451.2020.1734444

    • Related Report
      2019 Research-status Report
    • Peer Reviewed
  • [Journal Article] First synthesis and absolute configuration of phorbasin H, a diterpene carboxylic acid isolated from the sponge Phorbas gukulensis2019

    • Author(s)
      Takahashi, K., Ogura, Y., Kuse, M., Takikawa, H.
    • Journal Title

      Biosci. Biotechnol. Biochem.

      Volume: 83 Issue: 12 Pages: 2198-2201

    • DOI

      10.1080/09168451.2019.1654850

    • Related Report
      2019 Research-status Report
    • Peer Reviewed
  • [Journal Article] Enhanced elevations of hypo-osmotic shock-induced cytosolic and nucleic calcium concentrations in tobacco cells by pretreatment with dimethyl sulfoxide2019

    • Author(s)
      Nguyen HTH, Bouteau F, Mazars C, Kuse M, Kawano T
    • Journal Title

      Biosci Biotechnol Biochem

      Volume: 83 Issue: 2 Pages: 318-321

    • DOI

      10.1080/09168451.2018.1533801

    • Related Report
      2019 Research-status Report
    • Peer Reviewed / Int'l Joint Research
  • [Presentation] Pholasinの発現と活性化2021

    • Author(s)
      森口舞子、姜法雄、久世雅樹
    • Organizer
      日本農芸化学会 2021年度大会(仙台)
    • Related Report
      2020 Research-status Report
  • [Presentation] N-Ar-DCL誘導体の合成と発光活性2020

    • Author(s)
      森口舞子, 姜 法雄, 滝川浩郷, 久世雅樹
    • Organizer
      日本農芸化学会 2020年度大会
    • Related Report
      2019 Research-status Report
  • [Presentation] N, O-アリールデヒドロセレンテラジン誘導体の合成2019

    • Author(s)
      森口舞子, 姜 法雄, 滝川浩郷, 久世雅樹
    • Organizer
      日本農芸化学会 関西・中部支部 2019年度合同神戸大会
    • Related Report
      2019 Research-status Report
  • [Presentation] ヒカリカモメガイの光る仕組みを有機化学で解明する2019

    • Author(s)
      久世雅樹
    • Organizer
      日本農芸化学会中四国支部 第29回 若手シンポジウム
    • Related Report
      2019 Research-status Report
    • Invited
  • [Remarks] 神戸大学農学研究科 天然有機分子化学研究室

    • URL

      http://www.research.kobe-u.ac.jp/ans-nprd-chem/index.html

    • Related Report
      2021 Annual Research Report 2020 Research-status Report 2019 Research-status Report

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Published: 2019-04-18   Modified: 2023-01-30  

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