Figure 5.

JYQ-42 is a non-competitive inhibitor of SIRT6. (A) Competition assay between NAD⁺ and JYQ-42, as evaluated by the Fluor de Lys (FDL) assay. (B) Competition assay between Ac-RHKK-Ac-AMC and JYQ-42, as evaluated by the Fluor de Lys (FDL) assay. (C and D) Enzyme kinetics analysis of SIRT6 inhibition by JYQ-42 were performed with varying concentrations of NAD⁺(C) and Ac-RHKK-Ac-AMC (D) by HPLC. Double-reciprocal plots of the initial velocities (Lineweaver–Burk plots) showing non-competitive inhibition towards NAD⁺ and Ac-RHKK-Ac-AMC. Data are presented as the mean ± SD, n = 3 wells, from three independent experiments. (E) The sequence conservation of the allosteric pocket was analysed by using WebLogo⁶². The amino acids are coloured according to their chemical properties as follows: hydrophobic: black; hydrophilic: blue; neutral: green.