Stability of P450 2B6dH and mutants measured using different methods
Standard errors for fit to the respective equations are shown. Results are representative of at least two independent determinations. The variation between the experiments is ≤10%.
P450 | P450 (HS, LS, and P420) | Trp Fluorescence Tm,app | Activity T50 | H2O2 (60 mM) kinact | Heme Dissociation [GuHCl] | ||
|---|---|---|---|---|---|---|---|
| Tm | kinact | ||||||
| °C | min-1 | °C | °C | min-1 | M | ||
| M2B6dH | 48.2 ± 0.5a | 0.034 ± 0.01 | 51.1 ± 0.1 | 45.2 ± 0.3 | 0.025 ± 0.002 | 0.41 ± 0.02 | |
| M198L | 46.0 ± 0.4 | 0.125 ± 0.01 | N.D. | 41.5 ± 0.4 | 0.050 ± 0.003 | N.D. | |
| L264F | 52.2 ± 0.5 | 0.013 ± 0.01 | 52.6 ± 0.1 | 46.3 ± 0.2 | 0.022 ± 0.002 | 0.56 ± 0.02 | |
| L390P | 44.2 ± 05 | 0.157 ± 0.01 | 51.5 ± 0.2 | 41.8 ± 0.6 | 0.042 ± 0.004 | 0.30 ± 0.03 | |
N.D., Not determined; HS, high spin; LS, low spin