Table 1.
MitoA (10 μm) was incubated with each substance for the indicated time and temperature in KCl buffer and analyzed by RP-HPLC
Dimethyl trisulfide was dissolved in 70% EtOH. For NO, DetaNONOate (50 μm) was disolved in KCl buffer, which had been deoxygenated by bubbling with argon for 30 min. Superoxide was generated using 5 milliunits/ml of xanthine oxidase and 1 mm hypoxanthine in KCl buffer and its production quantified as the SOD-sensitive reduction of ferricytochrome c. Reactivity = (MitoN × 100)/(MitoN + MitoA).
| Concentration | Incubation condition | Reactivity | |
|---|---|---|---|
| % | |||
| H2S | 100 μm | 4 h, RTa | 94 |
| Glutathione | 5 mm | 24 h, 37 °C | 7.3 |
| 5 mm | 4 h, 37 °C | 2.2 | |
| 10 mm | 24 h, 37 °C | 7.9 | |
| 10 mm | 4 h, 37 °C | 4.6 | |
| Cysteine | 2 mm | 4 h, RT | <LODb |
| Dimethyl trisulfide | 100 μm | 4 h, RT | <LOD |
| Lipoic acid | 100 μm | 4 h, RT | <LOD |
| Dihydrolipoic acid | 100 μm | 4 h, RT | <LOD |
| Na2S2O3 | 100 μm | 4 h, RT | <LOD |
| Na2S2O4 | 100 μm | 4 h, RT | <LOD |
| NADH | 5 mm | 4 h, RT | <LOD |
| NADPH | 5 mm | 4 h, RT | <LOD |
| NaSCN | 100 μm | 4 h, RT | <LOD |
| NaNO2 | 100 μm | 4 h, RT | <LOD |
| NO | 50 μm | 4 h, RT | <LOD |
| ONOO− | 100 μm | 4 h, RT | 1.25 |
| H2O2 | 100 μm | 4 h, RT | <LOD |
| t-BuOOH | 100 μm | 4 h, RT | <LOD |
| O2˙̄ | 1.79 nmol of cytochrome c/min | 4 h, RT | <LOD |
| HOCl | 100 μm | 4 h, RT | <LOD |
a RT, room temperature.
b LOD, limit of detection.