Figure 3.

Purified recombinant SmNPP5 cleaves NAD. a. ε-NAD was incubated with 12.5, 25, or 50 ng of rSmNPP5, as indicated, and cleavage was assessed by an increase in fluorescence over time (n = 3, mean ± SEM) b. β-NAD was incubated with (+) or without (-) 1 µg rSmNPP5 for either 1 h or 5 h (as indicated) and phosphate levels (mean ±SEM) were assessed following CIP treatment. c. Thin Layer chromatography analysis of reaction products following NAD (black arrowhead) incubation with (+) or without (-) 1 µg rSmNPP5 for 1 h or 5 h (as indicated). Cleavage yields NMN (red arrowhead) and AMP (green arrowhead). Migration of standards is shown at left. NAD, nicotinamide adenine dinucleotide; NMN, nicotinamide mononucleotide; AMP, adenosine monophosphate. d. Depiction of NAD cleavage reaction catalyzed by SmNPP5; structures of NAD and cleavage products NMN and AMP are shown.