Fig. 3: Compound 15 is a covalent inhibitor of NEDD4.
From: Structure-based design of potent and selective inhibitors of the HECT ligase NEDD4
A The modification of Norclomipramine’s nitrogen methyl groups with various electrophilic warheads led to the discovery of compound (cmpd) 5, which exhibits low micromolar potency. B Close-up view of HECT NEDD4 bound to compound 15 (PDB 9H9T). Side chains of residues involved in inhibitor binding as well as residue Cys627, covalently bound to the inhibitor are highlighted. C Ub-TR-FRET assay performed with full-length protein NEDD4 pre-incubated for 1 hour with increasing concentrations of compound 15 before ATP addition and measurement. Plots show single time-point measurements of FRET signal to background ratio (% Signal/Background) as function of time (minutes). Mean ± SEM (n = 4). D Ub-TR-FRET assay as in C using the HECT domain of NEDD4. Mean ± SEM (n = 4).
