Table 1.
Ligand presence | Mock labeling reaction conditions | |||
---|---|---|---|---|
100 mM HEPES pH 7.5 | 100 mM HEPES pH 7.5, 20 % glycerol | 100 mM HEPES pH 7.5, 1 μM CPR-K56Q | 100 mM potassium phosphate, pH 7.4 | |
None | < 18 | 72 ± 6 | 120 ± 10 | 72 ± 6 |
+ 200 μM 7HC | < 18 | 100 ± 13 | 105 ± 18 | 79 ± 3 |
Experiments were performed in 100 mM potassium phosphate pH 7.4, 20 μM dilauroyl-α-L-phosphatidylcholine, 2 U μL−1 catalase, and 0.04 μg μL−1 superoxide dismutase, with 250 M pNP and a NADPH-regenerating system [2 μU μL−1 glucose 6-phosphate dehydrogenase, 10 mM glucose 6-phosphate, 2 mM MgCl2, 1 mM NADP+]. The final enzyme concentrations were 25 nM CYP2E1, 100 nM CPR-K56Q, and 50 nM cyt b5. The reported values reflect the results from average of 2–3 experiments including the standard deviation from the mean.