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

2017 Fiscal Year Final Research Report

• Project/Area Number: 26660289
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Applied molecular and cellular biology
• Research Institution: The 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
• 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.

Free Research Field

構造生物学、蛋白質工学