The physical biology of G protein-coupled receptor (GPCR) signalling can be inferred from imaging of single molecules and single living cells. In this opinion paper, we highlight recent developments in technologies to study GPCR signalling in vitro and in cyto. We start from mobility and localisation characteristics of single receptors in membranes. Subsequently, we discuss the kinetics of shifts in receptor-conformation equilibrium due to allosteric binding events and G protein activation. We continue with recent insights into downstream signalling and the role of delayed negative feedback to suppress GPCR signalling. Finally, we discuss new strategies to reveal how the multiplex signalling responses of cells to ligand mixtures, mediated by their entire receptor arsenal, can be disentangled, using single-cell data.

中文翻译：

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从荧光光谱和成像推断出GPCR信号动力学的物理生物学。

G蛋白偶联受体（GPCR）信号传导的物理生物学可以从单个分子和单个活细胞的成像中推断出来。在这篇意见书中，我们重点介绍了在体外和细胞中研究GPCR信号传导技术的最新进展。我们从膜中单个受体的迁移和定位特性开始。随后，我们讨论了由于构构结合事件和G蛋白活化导致的受体构象平衡偏移的动力学。我们继续对下游信号传导以及延迟负反馈抑制GPCR信号传导的作用进行了最新的了解。最后，我们讨论了新的策略，以揭示如何利用单细胞数据来理清细胞对配体混合物（由其整个受体阿森纳介导）的多重信号响应。