Properties of the nNOS inactivator(s) present in CE. The CE was treated under various conditions, as described under Materials and Methods. The untreated CE (None) or the various CE preparations (0.015 cigarette equivalents) were added to the first reaction mixture, and the activity of nNOS was determined after a 60-min incubation. The values are the mean ± S.E. (n = 3).
Treatment of CE | nNOS Activity Remaining at 60 Min |
|---|---|
| % Control | |
| None | 28.2 ± 2.3 |
| Evaporation by SpeedVac apparatus | 29.0 ± 3.6 |
| Ultrafiltration with a 3,000-Da molecular mass cut-off membrane | |
| Retentate | 102.3 ± 2.0 |
| Filtrate | 24.7 ± 4.3 |
| Methylene chloride extraction | |
| Fraction | |
| 1 | 86.3 ± 6.0 |
| 2 | 102.0 ± 4.2 |
| 3 | 86.7 ± 5.5 |
| 4 | 34.0 ± 4.6 |
| Binding to cation exchange columns | |
| Flow-through | 92.3 ± 6.7 |
| Eluent of 2 M NaCl | 26.0 ± 4.6 |
| Binding to anion exchange columns | |
| Flow-through | 18.0 ± 5.0 |
| Binding to activated charcoal | |
|
Supernatant after treatment
|
102.3 ± 3.7
|