Construction of ligand assay system for GABA(A) receptors using ligand-directed chemistry

2022 Fiscal Year Final Research Report

• Project/Area Number: 20K05746
• 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: Kyoto University
• Principal Investigator: Sakamoto Seiji 京都大学, 工学研究科, 特定准教授 (30335228)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Keywords: GABA(A)受容体 / リガンド指向性化学 / タンパク質ラベル化 / バイオセンサー / イメージングプローブ

Outline of Final Research Achievements

gamma-Aminobutyric acid (GABA) is the major inhibitory neurotransmitter in the central nervous system. The fast inhibitory actions of GABA are mainly mediated by GABA(A) receptors (GABA(A)Rs), which are widely recognized as clinically relevant drug targets. However, it remains difficult to create screening systems for drug candidates that act on GABA(A)Rs because of the existence of multiple ligand-binding sites and the delicate pentameric structures of GABA(A)Rs. In this study, we developed the first turn-on fluorescent imaging probe for GABAARs, which can be used to quantitatively evaluate ligand-receptor interactions under live cell conditions. In addition, with the aim of developing fluorescent biosensors using endogenous GABA(A)Rs in living mouse brains and neurons, we explored a new method based on ligand-directed chemistry that allow the chemical labeling of endogenous receptors without genetic manipulations.

Free Research Field

ケミカルバイオロジー

Academic Significance and Societal Importance of the Research Achievements

GABA(A) 受容体は神経機能制御に必須であり、その機能異常は不眠・不安・緊張・けいれん・てんかん等様々な病態を引き起こす事から、様々な神経疾患と精神疾患の治療標的にもなっている。従って、本研究で構築を試みる受容体型バイオセンサーは、脳高次機能に対する分子レベルでの基礎的知見を与えるのみならず、神経関連疾患に対する新規薬剤設計においても重要な指針を与えることが期待できる。さらに、生体分子間相互作用解析のために、高い認識能と感度を備えた機能分子センサーや新たな分析、計測手法の開発は必須であり、研究成果の技術的ニーズは高い。