Single-molecule imaging-based GPCRome analysis

2022 Fiscal Year Final Research Report

• Project/Area Number: 20K05760
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 37030:Chemical biology-related
• Research Institution: Institute of Physical and Chemical Research
• Principal Investigator: Yanagawa Masataka 国立研究開発法人理化学研究所, 開拓研究本部, 研究員 (70609792)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Keywords: GPCR / 1分子イメージング / 細胞内シグナル伝達 / アレスチン / エンドサイトーシス / ERKシグナル / 拡散機能相関

Outline of Final Research Achievements

G protein-coupled receptors (GPCRs) are major drug targets. In the present study, we analyzed the interaction between GPCRs and their effectors, including G protein, arrestin, and GRKs, at the single molecule level to elucidate the molecular basis that governs the diffusion dynamics of GPCRs in the plasma membrane. As a result, we found that arrestin binding is the main factor of the diffusion dynamics change that commonly observed in many GPCRs. We also revealed how GRK-dependent phosphorylation of GPCRs, which promotes arrestin binding, is regulated in model receptors. In addition, we developed the basic technology to perform comprehensive single-molecule imaging of GPCRs. We established a new fluorescent labeling method for GPCRs, which solved a problem in a conventional labeling method that caused unstable arrestin binding to a specific GPCR.

Free Research Field

生物物理学

Academic Significance and Societal Importance of the Research Achievements

GPCRが「いつ」「どこで」「どのように」エフェクター分子と相互作用し、細胞外から受容した情報を細胞内へと伝えるのかを解明することは、薬の作用機序を理解する上で重要な課題である。本研究では、GPCRとエフェクターの相互作用を1分子レベルで定量する手法を開発し、細胞内シグナル伝達の空間的な制御機構の一端を解明することに成功した。本研究で開発した細胞内1分子計測・解析のワークフローはGPCRだけでなく他の膜受容体を標的とした薬理学・創薬研究にも応用できると期待される。