Phase Separation Based Reporters of Protein-Protein Interactions

摘要

The invention is a novel reporter system for measuring protein-protein interactions. The system uses a pair of functionalized coiled coil subunits that spontaneously form two separate homo-oligomers when expressed in cells. The coiled coil subunits are functionalized with fluorescent proteins and complementary interacting proteins. Upon an activation stimulus which promotes the protein-protein interaction, the interacting proteins drive the formation of multivalent aggregates of the homo-oligomers in phase-shifted droplets. The highly concentrated fluorescent proteins in these structures provide high brightness over background fluorescence and a readily observed, quantitative and dynamic indicator of the protein-protein interaction. The reporters and assay methods are amenable to cells and whole organisms.