Disruption of NAD+ binding site in glyceraldehyde 3-phosphate dehydrogenase affects its intranuclear interactions

Figure 4 Kinetic analysis of wild type and mutated forms of Hisx6-tagged glyceraldehyde 3-phosphate dehydrogenase prepared in BL21 (DE3) Escherichia coli expression system. Conditions for the glycolytic assay are indicated in “Materials and Methods”. A: Reaction was performed with 0.26 mmol/L NAD⁺ and varying concentrations of D-glyceraldehyde-3-phosphate (0-2 mmol/L); B: Reaction was performed with 0.51 mmol/L D-glyceraldehyde-3-phosphate and varying concentrations of NAD⁺ (0-2 mmol/L). Vmax and Km were calculated by non-linear regression analysis with GraphPad Prizm 4.0 as described in Materials and Methods. GAPDH: Glyceraldehyde 3-phosphate dehydrogenase.