Table 3. Compactin Binding and Hydroxylation by P450 Variants.
| varianta | source | pravastatinb (%) | 6-epi-pravastatina (%) | Kd (μM) | LS/HSc conversion (%) |
|---|---|---|---|---|---|
| CYP105AS1 | wild type | 11 | 89 | 30.3 ± 5.2 | <5 |
| P450prac | error-prone PCR | 95 | 5 | 0.77 ± 0.07 | 70–80 |
| P450pra + F76N | rational | 85 | 15 | 4.81 ± 0.25 | 60–70 |
| P450pra + P80G | rational | 68 | 32 | 5.84 ± 0.34 | 70–80 |
| P450pra + T286I | rational | 71 | 29 | 4.93 ± 0.25 | 60–70 |
| P450pra + T95W | computational | 79 | 21 | 3.29 ± 0.03 | 60–70 |
| P450pra + T95F | computational | 95 | 5 | 0.25 ± 0.01 | >95 |
| P450pra +V180I | rational | 93 | 7 | 1.64 ± 0.13 | 80–90 |
| P450pra +V180M | computational | 97 | 3 | 1.34 ± 0.06 | >90 |
| P450pra + T95F/V180M (=P450pra100) | computational/rational | >99 | <1 | 0.13 ± 0.03 | >95 |
a
In comparison to wild type, P450pra carries the active site mutations I95T, A180V, and L236I, and on the surface (Q127R, A265N). The mutations above are on top of these mutations.
b
Percentage of total detected hydroxylation product.
c
Partial low-spin (LS, absorption at 418 nm) to high-spin (HS, at 390 nm) conversion was observed upon titration with compactin.