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. 2012 Jul 31;287(39):32525–32534. doi: 10.1074/jbc.M112.362715

FIGURE 2.

FIGURE 2.

NAADP is degraded to NAAD in HeLa cells. Cytosolic protein (S100), the light membrane fraction (P100), and the heavy membrane fraction (P10), each at 1 mg/ml, were incubated with 20 μm NAADP at 37 °C for 15 min. Reaction products were analyzed by HPLC. A, shown are representative chromatograms before and 15 min after incubation with substrate. Incubation of NAADP with membrane fractions from HeLa cells resulted in dephosphorylation of NAADP and formation of NAAD. B, the reaction product NAAD was quantified after 15 min of incubation. NAAD production by P10 membrane fractions was much higher than that by P100 membrane fractions. Incubation of NAADP with cytosolic fractions (S100) resulted in relatively little NAAD formation. Data shown are means ± S.D. (n = 7–14). **, p < 0.01; ***, p < 0.001 (analysis of variance).