Recent advances in the development of fluorescent ligands for G-protein-coupled receptors (GPCRs) and receptor tyrosine kinase receptors (RTKs) have facilitated the study of these receptors in living cells. A limitation of these ligands is potential uptake into cells and increased nonspecific binding. However, this can largely be overcome by using proximity approaches, such as bioluminescence resonance energy transfer (BRET), which localise the signal (within 10 nm) to the specific receptor target. The recent engineering of NanoLuc has resulted in a luciferase variant that is smaller and significantly brighter (up to tenfold) than existing variants. Here, we review the use of BRET from N-terminal NanoLuc-tagged GPCRs or a RTK to a receptor-bound fluorescent ligand to provide quantitative pharmacology of ligand–receptor interactions in living cells in real time.

G 蛋白偶联受体 (GPCR) 和受体酪氨酸激酶受体 (RTK) 荧光配体开发的最新进展促进了对活细胞中这些受体的研究。这些配体的一个局限性是可能被吸收到细胞中并增加非特异性结合。然而，这在很大程度上可以通过使用接近方法来克服，例如生物发光共振能量转移 (BRET)，它将信号（在 10 nm 内）定位到特定的受体目标。最近的 NanoLuc 工程已经产生了一种荧光素酶变体，它比现有变体更小但亮度明显更高（高达十倍）。这里，