Fig. 3: COOKIE-Pro profiling for spebrutinib in Ramos cell using SB-2 desthiobiotin probe.
From: COOKIE-Pro: covalent inhibitor binding kinetics profiling on the proteome scale
A Chemical structures of Spebrutinib (Speb) and the SB-2 probe. B Spebrutinib COOKIE experiment layout and the immunoblot for BTK in the pulldown samples. Blots are representative for three independent experiments. C Ranked proteomics -log2 transformed ACC1-normalized intensity ratio between 5 µM spebrutinib saturation sample and DMSO sample as an off-target protein list. D ACC1-normalized intensity ratio heatmaps for spebrutinib kinase off-targets across the COOKIE panel. E Representative first step progressive curve fitting for \({k}_{{{\rm{obs}}}}\) at different spebrutinib concentrations against BTK and TEC. F Second step [I]-kobs plots fitting into Michaelis–Menten model (MM-fitting) to obtain \({k}_{{{\rm{inact}}}}\) and \({K}_{I}\) values for BTK and TEC. Data are presented as mean ± SEM from three independent experiments. Source data are provided as a Source Data file.
