Role of the Substrate Specificity-Defining Residues of Human SIRT5 in Modulating the Structural Stability and Inhibitory Features of the Enzyme
Fig 10
ITC profile for the binding of EX527 to SIRT5 Y102A/R105I in the absence (left) and presence of 10 mM NAD+ (right).
The top panels show the raw calorimetric data generated by titration of 20 μM SIRT5 Y102A/R105I by 45 injections (5 μl each) of 500 μM EX527. The area under each peak was integrated and plotted against the molar ratio of EX527 to SIRT5 Y102A/R105I. The solid lines (right panel) represent the best fit of the experimental data for the single binding site model, yielding the magnitudes of n, ΔH° and Ka as being equal to 1.2, −5.3 ± 0.1 kcal/mol and (2.5 ± 0.2) × 105 M-1, respectively.