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HPLC analyses of the enzymatic products from NAD produced by CD38 and its mutants. WT CD38 (upper panel, 0.2 μg/ml), T221F (middle panel, 5 μg/ml), or T221F/E146A (lower panel, 2 μg/ml) was incubated with 100 μm NAD for the indicated times and analyzed by HPLC. There was no detectable cADPR production by WT CD38 under these conditions, the T221F mutant produced some detectable cADPR, and the T221F/E146A double mutant produced predominantly cADPR. Ab, absorbance.
