Author: Lee K, Navaluna E, Marsh M M, Janezic E M, Hague C.
SLAS Discovery, 2022
The authors of this study have developed a novel cyclo-heximide (CHX)-chase assay to monitor G protein-coupled receptor (GPCR) degradation in human cell lines. HEK293 cells were transfected with pSNAP and subsequently treated with CHX. The CytoSMART Exact (formerly known as the Corning Cell Counter) was used to count the cells after which they were diluted, lysed and the GPCR expression was determined via gel electrophoresis. This novel method was used to quantify the half-life of 21 different human GPCRs. In summary, the novel CHX-chase assay enables precise quantitative analysis of GPCR degradation in human cells.
