TABLE 6.
Profiles of two representative purifications of ovine complex I
NADH/FeCy activity was measured at each purification step.
| Volume | Protein | Total activity | Specific activity | |
|---|---|---|---|---|
| ml | mg | units | units/mg | |
| DDM purification | ||||
| Solubilized washed membranes | 53 | 584 | 3400 | 5.8 |
| Extract supernatant | 50 | 269 | 3200 | 11.9 |
| Q-Sepharose pooled fractions | 58 | 50 | 1900 | 38.0 |
| Superose 6 pooled fractions | 6 | 6.4 ± 0.6a | 530 ± 15a | 82.0 ± 7. b |
| LMNG purification | ||||
| Solubilized washed membranes | 55 | 514 | 3700 | 6.0 |
| Extract supernatant | 52 | 333 | 3000 | 7.2 |
| Q-Sepharose-pooled fractions | 45 | 61 | 860 | 15.7 |
| Superose 6-pooled fractions | 8 | 3.2 ± 0.2a | 230 ± 10a | 71.0 ± 4.8b |
a n = 3–5.
b The difference in the specific activity may stem from an overestimation of protein concentration due to LMNG carried over in the BCA assay as the presence of LMNG was not controlled in the BCA standards.