Figure 1.

Characterization of bovine GAPDH. A. SDS-PAGE analysis of bGAPDH purified from bovine ROS. B. Size exclusion profile of purified bGAPDH. The column (Superdex 200, 10/300 GL, GE Healthcare) was calibrated with protein standards (Insert). C. Enzyme activity. Purified bGAPDH (0.5 µg/100 µL) was incubated with 0.2 mM NAD and increasing concentrations of G3P (top panel), or with 1 mM G3P and increasing concentrations of NAD (middle panel). After 30 min of incubation, GAPDH activity was monitored by measuring NADH production at 340 nm. Kinetic measurements were then done over a 45 min time course in the presence of 1 mM NAD and 1 mM G3P (bottom panel).