Fig. 2: BCCov covalently labels BTK in cells.
From: A covalent BTK ternary complex compatible with targeted protein degradation
A Deconvoluted intact mass spectrum chromatograph of immunopurified BTKFL after DMSO treatment (red) or after cellular treatment with 20 µM BCCov (blue). Caliper indicates mass shift representing BCCov modification. Heterogeneous phosphorylation states 0P, 1P, or 2P represent 79.9 Da mass shifts. The inlay shows a Coomassie-stained gel showing Immunopurified BTKFL and corresponding far western blot with biotinylated-cIAP1BUCR1 detected with streptavidin HRP. Each is a representative image for 3 independent experiments. B, C Representative western blot for time-dependent degradation of BTKFL and vinculin loading controls after 5 µM BCCov or BCNC treatment, respectively. Each experiment was performed 3 times. D Graph of % covalent bond formation (red diamonds, right axis) versus observed BTKFL degradation (blue squares and black triangles for BCCov and BCNC respectively; left axis) over time. Each experiment was performed three times, with mean values plotted with SEM Error bars. Samples processed from different blots were from the same experiment and processed in parallel using DMSO treatment as a normalization control.
