Table 2.
Summary of yields of nitrite and GSNO from 1 µM DEA/NO in the absence and presence of 1 mM GSH observed with various assay methods.
| GSH | Mg2+ omitted | SOD omitted | KPi | Vessel closed | NO (µM) | NO2− (µM) | GSNO (µM) |
|---|---|---|---|---|---|---|---|
| − | I: 0.07±0.02 | ||||||
| − | II: 0.08±0.02 | II: 0.61±0.03 | |||||
| − | III: 0.60±0.05 | ||||||
| − | IV: 0.56±0.08 | ||||||
| − | V: 0.66±0.05 | ||||||
| − | VI: 0.43±0.02 | ||||||
| − | ⁎ | III: 0.37±0.06 | |||||
| − | ⁎ | V: 0.66±0.10 | |||||
| − | ⁎ | III: 0.48±0.05 | |||||
| − | ⁎ | III: 1.40±0.01 | |||||
| − | ⁎ | VI: 1.36±0.07 | |||||
| + | I: 0.02±0.01 | I: 0.94±0.05 | |||||
| + | II: 0.02±0.01 | II: 0.53±0.03 | II: 0.67±0.02 | ||||
| + | III: 0.53±0.05 | III: 0.69±0.04 | |||||
| + | IV: 0.71±0.10 | ||||||
| + | V: 0.74±0.09 | ||||||
| + | VI: 0.38±0.05 | VI: 1.20±0.20 | |||||
| + | ⁎ | I: 0.32±0.05 | |||||
| + | ⁎ | III: 0.36±0.01 | III: 0.25±0.01 | ||||
| + | ⁎ | V: 0.36 | |||||
| + | ⁎ | VI: 0.49±0.06 | |||||
| + | ⁎ | I: 0.42±0.06 | |||||
| + | ⁎ | VI: 0.56±0.07 | VI: 0.58±0.07 | ||||
| + | ⁎ | I: 0.51±0.01 | |||||
| + | ⁎ | III: 0.46±0.08 | III: 0.42±0.01 | ||||
| + | ⁎ | VI: 0.36±0.11 |
The following methods were used: NO electrode (method I), NO analyzer with KI/I2 and CuSO4 (method II), NO analyzer with sulfanilamide and KI/I2 (method III), fluorimeter with DAN (method IV), fluorimeter with commercial nitrite/nitrate kit (method V), and HPLC (method VI). See Materials and methods for further details. Experimental conditions: 1 µM DEA/NO, 1 mM GSH as indicated, 1000 U/ml SOD as indicated, 0.1 mM DTPA, and 5 mM MgCl2 as indicated in 50 mM triethanolamine–HCl (or 50 mM KPi) buffer (pH 7.4) in 0.5 ml at 37 °C; yields were determined after 12 min incubation in open or closed vessels as indicated.