Bridging the gap between single molecule and bulk fluorescence techniques with PIE-FCCS

Abstract

G protein-coupled receptors (GPCRs) are a diverse set of transmembrane proteins with large variations in sequence and domain topology. One important aspect of GPCR function is the role of spatial organization and dimerization/oligomerization, but there is still no systematic understanding of these effects. This is due, in part, to the techniques that have been used to analyze them. Bulk fluorescence techniques cannot fully quantify receptor concentration, diffusion and oligomerization state. Single molecule tracking techniques are excellent at determining membrane protein diffusion and protein-protein interactions, but the technology requires very low expression levels and so is challenging to perform at physiological expression levels.