The –SH Protection Method for Determining Accurate Kd Values for Enzyme-Coenzyme Complexes of NAD+-Dependent Glutamate Dehydrogenase and Engineered Mutants: Evidence for Nonproductive NADPH Complexes
Figure 2
Protection by NADPH against inactivation by DTNB. The plots show for wild-type GDH and three mutants the dependence on NADPH concentration of the pseudo-first-order rate constant for inactivation of the enzyme by 200 μM DTNB under the same conditions as in Figure 1. The generalised equation giving rise to this plot is (3), in this case being . The inset plot shows data for wild-type GDH. In the main plot the data are for F238S (squares), P262S (triangles, upper line), and F2238S/P262S (inverted triangles).
