使用 IP 均相时间分辨荧光（HTRF）测定法定量 Gq 信号转导。
Quantifying Gq Signaling Using the IP Homogenous Time-Resolved Fluorescence (HTRF) Assay.
机构信息
Metabolism and Systems Science, University of Birmingham, Birmingham, UK.
Centre of Membrane Proteins and Receptors (COMPARE), Universities of Birmingham and Nottingham, Birmingham, UK.
出版信息
Methods Mol Biol. 2025;2861:23-32. doi: 10.1007/978-1-0716-4164-4_2.
G protein-coupled receptors (GPCRs) play pivotal roles in cellular signaling and can regulate several cellular functions such as proliferation, secretion, protein expression, and cellular metabolism. Coupling of GPCRs to members of the Gq/11 protein family results in activation of inositol trisphosphate (IP3) and accumulation of calcium intracellularly. This protocol chapter outlines a step-by-step guide for utilizing the inositol phosphate-1 (IP) accumulation assay, a time-resolved fluorescence resonance energy transfer (TR-FRET) method, to investigate Gq-IP3 signaling. The assay serves as a valuable tool for those conducting pharmacological investigations and compound screening targeting this critical cellular pathway. This protocol chapter covers experimental setup, sample preparation, and data analysis, providing researchers with an in-depth guide to explore the pharmacology of Gq-coupled receptors.
G 蛋白偶联受体（GPCRs）在细胞信号转导中发挥着关键作用，能够调节多种细胞功能，如增殖、分泌、蛋白质表达和细胞代谢。GPCR 与 Gq/11 蛋白家族成员偶联会导致三磷酸肌醇（IP3）的激活和细胞内钙的积累。本方案章节概述了使用肌醇磷酸盐-1（IP）积累测定法（一种时间分辨荧光共振能量转移（TR-FRET）方法）研究 Gq-IP3 信号转导的分步指南。该测定法是针对该关键细胞途径进行药理学研究和化合物筛选的有力工具。本方案章节涵盖了实验设置、样品制备和数据分析，为研究人员提供了深入探索 Gq 偶联受体药理学的指南。
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