Steady-state kinetics: CuOOH-supported oxidation of 7-BQ by CYP3A4 and site-directed and random mutants in the presence of cytochrome b5
Results are representative of at least two independent determinations. The variation between the experiments is ≤20%.
CYP3A4 | kcat | Km, CuOOH | kcat/Km |
|---|---|---|---|
| min-1 | mM | ||
| WT | 3.9 (0.23)a | 0.54 (0.05) | 7.2 |
| L216W | 4.1 (0.29) | 0.31 (0.04) | 13 |
| F228I | 4.0 (0.16) | 0.41 (0.04) | 9.7 |
| T433S | 6.0 (0.31) | 0.49 (0.06) | 7.6 |
| T309A | 9.0 (0.46) | 0.52 (0.05) | 17 |
| T309V | 11 (0.45) | 1.2 (0.17) | 9.1 |
| F228I/T309A | 18.0 (0.56) | 0.60 (0.05) | 30 |
↵ a Standard errors for fit to Michaelis-Menten are shown in parentheses.