Development of site-specific labeling method of single-transmembrane receptors

2019 Fiscal Year Final Research Report

• Project/Area Number: 17K19206
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Research Field: Biomolecular chemistry and related fields
• Research Institution: Yokohama City University
• Principal Investigator: Nogi Terukazu 横浜市立大学, 生命医科学研究科, 准教授 (40379102)
• Project Period (FY): 2017-06-30 – 2020-03-31
• Keywords: １回膜貫通型受容体 / 半合成 / 受容体活性化機構 / コンフォメーション変化

Outline of Final Research Achievements

In this project, we have attempted to develop a novel technique to introduce a site-specific label into single-transmembrane receptors expressed on the cell membrane of higher organisms such as human. The site-specific labeling method will be utilized to detect the conformational change of the single-transmembrane receptor, which is induced from the interactions with extracellular signaling proteins. We have produced extracellular domain fragments of the single-transmembrane receptors with post-translational modifications such as glycosylation and disulfide bonds by using mammalian expression systems. We have tested the ligation between the extracellular domain fragments and peptides and optimized the reaction conditions. Furthermore, we have examined the influence of the solution conditions on the conformational change of the single-transmembrane receptors through the dynamic structural analysis methods.

Free Research Field

構造生物学

Academic Significance and Societal Importance of the Research Achievements

生体を構成する細胞は、細胞膜上に受容体タンパク質を配して外的環境からのシグナルを受容し、細胞内へと情報を受け渡す。この細胞表面受容体によるシグナル受容と変換は、分裂、分化、移動などの細胞の運命を決定する出発点とも言える。また変異等による受容体の機能異常は、がんを代表例とする様々な疾患を引き起こす。したがって、本研究で取り組んだ細胞表面受容体によるシグナル伝達の分子機構の解明は、細胞挙動の制御による疾患の治療に向けた新規な創薬戦略の提案にもつなる可能性があるという点において重要な意義をもつ。