Fig. 2: BRET probe CBH-002 enables live-cell target engagement assay for all three subclasses of PRMT5 inhibitors.
From: A BRET biosensor for measuring uncompetitive engagement of PRMT5 complexes in cells
A Affinity curve for BRET probe CBH-002 in HEK293 cells transfected with NanoLuc-PRMT5 and unlabeled WDR77 DNA. CBH-002 was titrated against 10 µM GSK3326595 or DMSO, and apparent affinity (EC50) was calculated from a sigmoidal 4-parameter curve-fit (see “Methods”). Data are the mean ± SEM of 3 independent experiments (n = 3). B Optimization of BRET probe concentration for live-cell TE. An 8 pt, 2-fold series of CBH-002 and an 11 pt, 3-fold dilution series of GSK3326595 were added to HEK293 cells expressing NanoLuc-PRMT5 fusion DNA and unlabeled WDR77 DNA. Optimal BRET probe concentration shown in orange. Data are the mean ± SEM of 3 independent experiments (n = 3). C–E Schematic representations of the three major PRMT5 inhibitor subclasses (top) with structures of representative compounds (middle), and live-cell NanoBRET data (bottom). Data are the mean ± SEM of 3 independent experiments (n = 3). Source data are provided as a Source Data file.
