Comparison of Methods used to Measure NADH-Dependent GSNO-R Activity.

Lungs were harvested from adult (10–12 w) C57Bl6/129SeEv mice. Lung homogenates were subjected to NADH-dependent GSNO-R activity. The assay was performed as described in Methods. The effect of altering the concentration of GSNO in the assay and the precipitation of protein prior to measurement were compared in the same samples (n = 4). Note that higher activity was obtained in the absence of protein precipitation for both GSNO concentrations. Highest GSNO-R activity was obtained in the presence of 100 µM GSNO. GSNO-R activity was found to be 2–3 fold higher in the lung homogenates from female animals regardless of the method used to measure activity.实验取材自10~12周龄成年C57Bl6/129SeEv小鼠的肺部组织。对肺组织匀浆开展还原型烟酰胺腺嘌呤二核苷酸（NADH）依赖型GSNO还原酶（GSNO-R）活性检测，实验操作参照本研究方法部分所述流程进行。针对同一份样本（n=4），分别比较了实验体系中调整GSNO浓度与检测前进行蛋白沉淀对检测结果的影响。需注意：两种GSNO浓度下，未进行蛋白沉淀时均可获得更高的酶活性；当GSNO浓度为100 μM时，可检测到最高的GSNO-R活性。无论采用何种检测方法，雌性小鼠肺组织匀浆中的GSNO-R活性均较雄性高出2~3倍。