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. Author manuscript; available in PMC: 2009 Dec 15.
Published in final edited form as: Arch Biochem Biophys. 2008 Sep 10;480(2):132–137. doi: 10.1016/j.abb.2008.09.001

Figure 3.

Figure 3

Activities of purified Nor1p(48-450). (A) Time course of the absorbance change of NADH (~0.13 mM) at 340 nm in the presence of saturated NO. The hatch mark indicates the time when enzyme (~130 nM) was added to the sample. There was no oxidation observed with NADPH. (B) Time course change of NO peak area measured by NOA. DEA-NONOate (1mM) was allowed to react in 100 mM phosphate buffer, pH 7.5, 5 mM NADH for 30 min at 37°C. The reaction sample (2 mL in a 5 mL gas-tight vial) was then cooled to 25°C before enzyme (~130 nM) was added. Headspace (15 μL) was sampled every few minutes. In the absence of enzyme, there was no significant change in the NO concentration (data not shown). (C) Time course change of N2O peak area measured by GC-ECD. DEA-NONOate (500 μM) was allowed to react with 2 mL NADH/phosphate buffer in a 15 mL gas-tight vial before enzyme (~40 nM) was added to initiate the reaction. Headspace (5 μL) was sampled every few minutes. Each time point represents duplicate samples and the error bars represent ranges.