TABLE 3.
Apparent KM and kcat values of CYP2E1 WT for CPR as measured by p-NP hydroxylation in the absence and presence of CYP2B4
CYP2E1 (0.1 μM) was reconstituted with varying concentrations of CPR in the presence or absence of 0.1 μM CYP2B4. The reconstituted mixtures were then added to 50 mM potassium phosphate buffer, pH 7.4, containing 0.3 mM p-NP, and 2 mM ascorbic acid. The reactions were initiated by adding 0.4 mM NADPH, and the incubations and determination of 4-NC formation were performed as described under Materials and Methods. The kinetic values given here were derived from data as plotted in Fig. 8.
| CYP2E1 WT |
||
|---|---|---|
| Control | Plus CYP2B4 | |
| KM (μM) | 0.14 ± 0.020 | 0.045 ± 0.01 |
| kcat (min–1) | 4.2 ± 0.015 | 5.65 ± 0.29 |