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Studies on the Construction of Biocatalysts with the Structural Framework of GPCR

Research Project

Project/Area Number 26660289
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Applied molecular and cellular biology
Research InstitutionThe University of Tokyo

Principal Investigator

ARAKAWA Takatoshi  東京大学, 大学院農学生命科学研究科(農学部), 助教 (30523766)

Project Period (FY) 2014-04-01 – 2018-03-31
Project Status Completed (Fiscal Year 2017)
Budget Amount *help
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2016: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000)
Fiscal Year 2015: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2014: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Keywordsタンパク質工学 / 代謝型受容体 / 膜タンパク質
Outline of Final Research Achievements

Growing number of structures are available for the integral membrane proteins. Especially G protein coupled receptors (GPCRs) are the major family encoded by mammalian genomes and extracellular surfaces of them are diversified and are specific for the respective ligands. While several bacterial and algal paralog proteins of GPCR have the other functions such as channels or pumps, no proteins are found as a seven transmembrane (7-TM) enzyme. Here our team performed first attempts to adapt the 7-TM frameworks to enzymes. Point mutations are done for the two residues of the agonist-binding site of the A2a adenosine receptor and then subtle changes in the affinities for the agonist are observed compared with the intact protein. Together, efforts are also focused on the preparation of GPCRs to visualize the resultant 7-TM catalysts. Through the trials of four other class A GPCRs, we have developed the methodology for large-scale preparations, stability analyses, and crystallizations.

Report

(5 results)
  • 2017 Annual Research Report   Final Research Report ( PDF )
  • 2016 Research-status Report
  • 2015 Research-status Report
  • 2014 Research-status Report
  • Research Products

    (1 results)

All 2017

All Presentation (1 results)

  • [Presentation] ドーパミン受容体の結晶化に向けた発現と精製2017

    • Author(s)
      和田京子、Yuwono Wibowo、伏信進矢、荒川孝俊
    • Organizer
      2017年度生命科学系学会合同年次大会
    • Related Report
      2017 Annual Research Report

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Published: 2014-04-04   Modified: 2019-03-29  

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