Bimolecular fluorescence complementation: lighting up seven transmembrane domain receptor signalling networks

摘要: There is increasing complexity in the organization of seven transmembrane domain (7TM) receptor signalling pathways, and ability their ligands to modulate direct this signalling. Underlying these events a network protein interactions between 7TM receptors themselves associated effectors, such as G proteins β-arrestins. Bimolecular fluorescence complementation, or BiFC, technique capable detecting protein–protein essential for function. Fluorescent proteins, those from Aequorea victoria, are split into two non-fluorescent halves, which then tag under study. On association, fragments refold regenerate mature fluorescent protein, producing BiFC signal indicative complex formation. Here, we review experimental criteria successful application considered context dimerization, β-arrestin The advantages limitations imaging compared with alternative resonance energy transfer techniques. We show that simplicity measurement allows high-content advanced applications, it can probe more multi-protein alone combination transfer. These capabilities suggest techniques will become ever useful analysis ligand pharmacology at molecular level interactions. This article part themed section on Imaging Pharmacology. To view editorial visit http://dx.doi.org/10.1111/j.1476-5381.2010.00685.x