Deconvolution of Complex G Protein-coupled Receptor Signaling in Live Cells: Taking Advantage of the Resolving Power of Dynamic Mass Redistribution Technology

Dynamic mass redistribution (DMR) represents a novel, label-free, biosensor technology promising to translate GPCR signaling into complex optical “fingerprints” in real time and living cells. Herein, we present a strategy to frame cellular mechanisms that define label-free responses and compare DMR technology with traditional second messenger assays currently state of the art in GPCR drug discovery. The resolving power of DMR enabled us to (1) probe GPCR functionality along all four G protein signaling pathways – at present beyond reach by many other traditional GPCR assay platforms, (2) dissect complex GPCR signaling patterns even in primary human cells with unprecedented accuracy, (3) define heterotrimeric G proteins as the upstream trigger orchestrating the complex optical response profiles, and (4) disclose previously undetected features of GPCR behavior. Significant impact of DMR is therefore anticipated in the emerging areas of systems biology and systems pharmacology but also for the discovery of mechanistically novel drugs.