Figure 5.

Dut80α D95E mutation decrease protein activity and stability. (A) dUTPase activity of Dut80α WT (blue circles) and D95E mutant (orange squares) was measured by malachite green assays. The initial velocities were calculated at different dUTP concentrations and kinetics parameters (Table 3) were determined from these curves using GraphPad software. The curves are the result of six independent assays. (B) Thermal-shift assays. Representative thermal denaturation curve profiles of Dut80α wt and D95E mutant in its apo states and in presence of MgCl₂ or dUPNNP-MgCl₂. The calculated Tm from at least three independent experiments are shown in Table 3.