Addition to the Primary Reaction Mixtures | Turnover Number | Activity Remaining |
---|---|---|
min−1 | % | |
Before gel filtration | ||
Water | 54 ± 1 3-a | 100 |
NADPH | 37 ± 3 | 68 |
After gel filtration | ||
Water | 50 ± 8 | 92 |
NADPH | 34 ± 4 | 62 |
The P-450 reconstituted system containing CYP2B1 and reductase was incubated for 10 min at 37°C with KR-II (50 μM) and either NADPH (1 mM) or water as described in Materials and Methods. Aliquots were then taken either for immediate analysis or for gel filtration with subsequent analysis of aliquots for activity in the benzphetamineN-demethylase assay.
↵3-a Mean ± S.D., n = 3.