G PROTEIN-COUPLED RECEPTOR (GPCR) LIGAND ASSAY

摘要

The systems and methods described herein include the use of Giant Plasma Membrane Vesicles (GPMVs) derived from cells expressing SPASM sensors (e.g., SPASM GPCR sensors). Herein provided are giant plasma membrane vesicle (GPMV) sensors containing a membrane-associated polypeptide, where the membrane-associated polypeptide contains a target G protein-coupled receptor (GPCR), a linker including an amino acid sequence flanked by a FRET donor and a FRET acceptor, and a receptor partner for the target GPCR, where the linker separates the target GPCR from the receptor partner. In another aspect, provided is a method for quantifying the intrinsic efficacy of a compound for a target GPCR. In another aspect, provided is a method for identifying a compound that interacts with a target GPCR.