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Construction of assay system for the ligand of G protein-coupled receptor using a fission yeast

Research Project

Project/Area Number 25650032
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Functional biochemistry
Research InstitutionTokyo Institute of Technology

Principal Investigator

Toshiya Osada  東京工業大学, 生命理工学研究科, 准教授 (00201997)

Project Period (FY) 2013-04-01 – 2016-03-31
Project Status Completed (Fiscal Year 2015)
Budget Amount *help
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2015: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2014: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2013: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
KeywordsGPCR / fission yeast / pheromone / GFP / フェロモン / グルコース / 分裂酵母 / リガンドセンサー / 創薬
Outline of Final Research Achievements

G-protein-coupled receptors (GPCRs) are seven transmembrane proteins and activated by hormones, odorants, neurotransmitters, and so on. GPCRs are one of the most potential drug targets. However, it is difficult to reproduce functional GPCR expression in heterologous cells due to the unknown mechanism of its trafficking and folding. The purpose of this project is the development of the ligand screening system for orphan GPCR and the construction of biosensor that utilizes the GPCR. The reporter strains for detecting a ligand response for an exogenous GPCR were constructed by using fission yeast. The fission yeast has two GPCR pathway. One is the glucose pathway and the other is the pheromone pathway. In this project, GFP reporter systems for both receptors have been constructed. The both transformed cells showed the ligand dependence activity. To increase the expression of the exogenous GPCR in the cells, the exogenous GPCR was modified with P3 signal derived from the fission yeast.

Report

(4 results)
  • 2015 Annual Research Report   Final Research Report ( PDF )
  • 2014 Research-status Report
  • 2013 Research-status Report
  • Research Products

    (11 results)

All 2016 2015 2014 2013

All Journal Article (3 results) (of which Peer Reviewed: 3 results) Presentation (8 results)

  • [Journal Article] Spectrin-ankyrin interaction mechanics: A key force balance factor in the red blood cell membrane skeleton2015

    • Author(s)
      Masakazu Saito, Takahiro Watanabe-Nakayama, Shinichi Machida, Toshiya Osada, Rehana Afrin, Atsushi Ikai
    • Journal Title

      Biophysical Chemistry

      Volume: 200-201 Pages: 1-8

    • DOI

      10.1016/j.bpc.2015.03.007

    • Related Report
      2015 Annual Research Report 2014 Research-status Report
    • Peer Reviewed
  • [Journal Article] Interaction between peptide pheromone or its truncated derivatives and pheromone receptor of the fission yeast Schizosaccharomyces pombe examined by a force spectroscopy study and a GFP reporter assay2013

    • Author(s)
      Hidaka S, Nikaido O, Kiyosaki S, Ikai A, and Osada T
    • Journal Title

      Journal of Surface Engineered Materials and Advanced Technology

      Volume: 3 Pages: 36-42

    • Related Report
      2013 Research-status Report
    • Peer Reviewed
  • [Journal Article] Gene expression of bovine vomeronasal receptors2013

    • Author(s)
      Sasuga S, Hidaka S, Ohkura S, Okamura H, and Osada T
    • Journal Title

      EJBS

      Volume: 7-05

    • Related Report
      2013 Research-status Report
    • Peer Reviewed
  • [Presentation] 分裂酵母内での外来性GPCRの発現2016

    • Author(s)
      岸本祐樹、長田俊哉
    • Organizer
      第10回日本ゲノム微生物学会年会
    • Place of Presentation
      東京
    • Year and Date
      2016-03-04
    • Related Report
      2015 Annual Research Report
  • [Presentation] 分裂酵母におけてグルコースシグナル経路のGαサブユニットであるGpa2発現レベルはフェロモンシグナル経路のシグナル伝達に影響を与える2015

    • Author(s)
      日高翔、長田俊哉
    • Organizer
      第38回日本分子生物学会年会
    • Place of Presentation
      神戸
    • Year and Date
      2015-12-01
    • Related Report
      2015 Annual Research Report
  • [Presentation] 分裂酵母における2種類のGPCRシグナルトランスダクション系間のクロストーク2014

    • Author(s)
      日高 翔、長田 俊哉
    • Organizer
      日本分子生物学会年会
    • Place of Presentation
      横浜
    • Year and Date
      2014-11-25 – 2014-11-27
    • Related Report
      2014 Research-status Report
  • [Presentation] 分裂酵母GPCR-Rgs1融合タンパク質の活性評価2014

    • Author(s)
      大屋 昌士、長田 俊哉
    • Organizer
      日本分子生物学会年会
    • Place of Presentation
      横浜
    • Year and Date
      2014-11-25 – 2014-11-27
    • Related Report
      2014 Research-status Report
  • [Presentation] 分裂酵母フェロモン受容体のN末端領域の機能解析2013

    • Author(s)
      有本 航、長田俊哉
    • Organizer
      日本分子生物学会年会
    • Place of Presentation
      神戸
    • Related Report
      2013 Research-status Report
  • [Presentation] 分裂酵母Gタンパク質Gpa1、Gpa2のN末端配列欠損が各シグナル伝達に与える影響2013

    • Author(s)
      小久保悟 長田俊哉
    • Organizer
      日本分子生物学会年会
    • Place of Presentation
      神戸
    • Related Report
      2013 Research-status Report
  • [Presentation] 分裂酵母Gタンパク質C末端の機能解析2013

    • Author(s)
      日高翔 長田俊哉
    • Organizer
      日本分子生物学会年会
    • Place of Presentation
      神戸
    • Related Report
      2013 Research-status Report
  • [Presentation] 分裂酵母フェロモン受容体の細胞内ループ置換がレポーターアッセイに与える影響2013

    • Author(s)
      小久保悟、長田俊哉
    • Organizer
      日本生化学会
    • Place of Presentation
      横浜
    • Related Report
      2013 Research-status Report

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Published: 2014-07-25   Modified: 2019-07-29  

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