Fig. 5: The recruitment of FBXO22 by G-6599 involves FBXO22 cysteine modification.
From: Rational design of potent small-molecule SMARCA2/A4 degraders acting via the recruitment of FBXO22
a Correlation between SMARCA2 degradation potency (DC50) and in vitro cysteine reactivity half-life for a series heteroaryl alkyne monovalent degraders. b Intact mass spectrometry of 1 µM of FBXO22/SKP1 alone (top) or in the presence of 20 µM of G-6599 with (middle) or without (bottom) 1 µM of SMARCA2 bromodomain. c Identification of covalently modified FBXO22 peptides using mass spectrometry after 4 h treatment of 1 µM FBXO22/SKP1 and SMARCA2 bromodomain in the presence of 50 µM of G-6599 for Cys228 (top) and Cys326 (bottom) modification. d Effect of varying concentrations of G-6599 on SMARCA2 levels in FBXO22 knockout HCC515 cells reconstituted with V5-tagged wild-type or mutated forms of FBXO22. Nuclear intensity of SMARCA2 within V5-positive cells is presented relative to DMSO control conditions. Data are presented as mean ± sd, n = 3 independent replicates. e anti-V5 immunoblot from lysates of HCC515 cells expressing V5-tagged FBXO22 mutants treated with G-6599 or DMSO control after exposure to a 54–64 °C temperature range. Actin served as a loading control. Source data are provided as a Source Data file.
