Steady-state kinetics: CuOOH-supported oxidation of 7-BQ by CYP3A4 and site-directed and random mutants
Results are representative of at least two independent determinations. The variation between the experiments is ≤15%.
CYP3A4 | kcat | Km, CuOOH | kcat/Km |
|---|---|---|---|
| min-1 | mM | ||
| WT | 1.2 (0.06)a | 1.2 (0.10) | 1.0 |
| T309A | 6.0 (1.0) | 1.2 (0.14) | 5.0 |
| T309V | 8.9 (1.5) | 1.4 (0.12) | 6.4 |
| L216W | 1.4 (0.12) | 0.44 (0.12) | 3.2 |
| F241V | 2.2 (0.09) | 1.0 (0.12) | 2.2 |
| F228I | 3.0 (0.16) | 0.70 (0.11) | 4.3 |
| T433S | 2.5 (0.09) | 1.3 (0.13) | 1.9 |
| L216W/F241V | 1.8 (0.10) | 1.4 (0.2) | 1.3 |
| F228I/T433S | 1.3 (0.09) | 1.0 (0.19) | 1.3 |
| L216W/F228I | 2.7 (0.20) | 1.8 (0.38) | 1.5 |
| F241V/F228I | 4.5 (0.29) | 2.6 (0.35) | 1.7 |
| F241V/T433S | 3.2 (0.15) | 2.3 (0.24) | 1.4 |
| L216W/F228I/T433S | 2.5 (0.26) | 1.2 (0.30) | 2.1 |
| L216W/T309A | 5.6 (0.29) | 0.70 (0.07) | 8.0 |
| F228I/T309A | 7.8 (0.50) | 0.73 (0.15) | 11 |
↵ a Standard errors for fit to Michaelis-Menten are shown in parentheses.