Extended Data Fig. 6: BLUE compounds are allosteric inhibitors and selective for human BRISC.
From: Molecular glues that inhibit deubiquitylase activity and inflammatory signaling
a, Chemical structure of JMS-175-2 and analogues FX-25-1, FX-25-2, which have substitutions in the piperidine ring (highlighted in green). b, Dose-response inhibition of BRISC by indicated compounds. IC50 values: JMS-175-2 = 3.8 µM, FX-25-1 = 5.2 µM, FX-25-2 = 21 µM. Data points are mean ± SEM of three independent experiments carried out in technical duplicate. c, Multiple sequence alignment (black = conserved, white = not conserved) of Abraxas1 and Abraxas2 from indicated species. Coloured boxes indicate BLUE interacting residues. d, FX-171-C inhibition of different BRISC orthologues. Hs - H. sapiens, Mm - M. musculus, Dr - D. rerio, Cf - C. floridanus. Data points are mean ± SEM of three independent experiments carried out in technical duplicate. e, f, Multiple sequence alignment of e, BRCC36 and f, BRCC45 from indicated BRISC orthologues. Residues are coloured as in c. g, Mass photometry analyses of dimer formation with FX-171-C for HsBRISCΔNΔC and HsBRISCΔLoop. Fraction of counts corresponding to BRISC dimer are plotted. Data points are mean ± SEM of three independent experiments. h, Negative stain EM 2D class averages of HsBRISCΔLoop incubated with FX-171-C. 22% of particles in the 2D class averages correspond to BRISC dimers. i, BRISC-FX-171-C cryo-EM density map highlighting an extended loop in BRCC36 (dust cleaning size 7.1, map threshold 0.0044).
