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. 2023 Jul 5;62(14):2147–2160. doi: 10.1021/acs.biochem.2c00599

Figure 3.

Figure 3

Evidence for covalent modification of WRN by 2-sulfonamide pyrimidines. The top panels show real-time DNA unwinding progress curves with varying concentrations of H3B-219 (A) and H3B-960 (B). Assays contained 0.6 nM WRN helicase domain, 0.5 μM Hel-10bp, 5 μM Trap-10bp, 120 μM ATP, and the indicated compound concentrations. The bottom panels show the intact mass spectra of WRN helicase domain (expected MW = 50939.5 Da) treated overnight with 10-fold excess of H3B-219 (C) or H3B-960 (D). Alkylation by H3B-219 and H3B-960 is expected to shift the mass by 253 or 307 Da adduct, respectively. For H3B-960, the indicated masses at 1× and 2× correspond to the mono- and dialkylated protein.