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Covalent inactivation of WRN helicase domain. (A) Proposed two-step mechanism where the initial E–I* complex is a reversible binding event (KI) prior to the inactivation step that forms the covalent E–I complex (kinact). The compounds H3B-859 (B), H3B-219 (C), and H3B-960 (D) follow this two-step mechanism. Real-time DNA unwinding assays were run with 0.5 nM WRN helicase domain, 120 μM ATP, 500 nM Hel-10bp, and 5 μM Trap-10bp.
