G-protein coupled receptors signaling; opsin; luciferase-based assays

Mutations in G-protein coupled receptors (GPCRs) affecting their function and signaling capabilities were shown to be associated with many human diseases. GPCRs are thus one of the most promising therapeutic targets. Establishment of assays enabling high throughput screenings of GPCR signaling are therefore highly warranted. In this review we provide overview of new strategies used for examination of GPCR signaling cascades. We focus mainly on recent studies using modified cyclic AMP sensitive luciferase enzyme. These assays enable detection of GPCR activation in living cells in real time. Moreover, this experimental design allows simultaneous and repeated measuring of GPCR association with any of possible downstream signaling cascades in one experimental setup. We summarize also papers presenting utilization of luciferase-based assays for analysis of opsins, light sensitive GPCRs, whose activation occurs in range of milliseconds.