TABLE 4.
Binding kinetics of RPV to WT and mutant enzymes
Values in parenthesis represent -fold change with respect to the WT enzyme.
| Enzyme | Kd.RPVa | kon.RPVa | koff.RPVa | Kd.RPVb | kon.RPVb | koff.RPVb |
|---|---|---|---|---|---|---|
| nm | nm−1 s−1 | s−1 | nm | nm−1 s−1 | s−1 | |
| WT | 17.9 ± 2.1 (1) | 0.07 ± 0.01 (1) | 1.3 ± 0.2 (1) | 13.9 ± 1.7 (1) | 0.14 ± 0.03 (1) | 1.9 ± 0.3 (1) |
| p66M184I/p51WT | 21.1 ± 1.6 (1.2) | 0.06 ± 0.01 (0.9) | 1.3 ± 0.1 (0.9) | 10.5 ± 1.1 (0.8) | 0.18 ± 0.02 (1.3) | 1.9 ± 0.3 (1) |
| p66E138K/p51E138K | 31.9 ± 1.9 (1.8) | 0.33 ± 0.02 (4.7) | 9.0 ± 0.8 (7.2) | 30.9 ± 2.3 (2.2) | 0.32 ± 0.03 (2.3) | 11.5 ± 1.8 (5.9) |
| p66E138K/M184I/p51E138K | 36.8 ± 1.3 (2.1) | 0.15 ± 0.02 (2.1) | 5.3 ± 0.6 (4.2) | 26.8 ± 1.9 (1.9) | 0.28 ± 0.01 (2.0) | 7.5 ± 1.1 (3.9) |
| p66M184I/p51E138K | 28.4 ± 2.3 (1.6) | 0.31 ± 0.01 (4.4) | 5.3 ± 0.6 (4.2) | 32.9 ± 2.6 (2.4) | 0.29 ± 0.05 (2.1) | 9.5 ± 1.2 (4.9) |
a Values obtained by fitting data to Equations 4–6. Also, Kd.RPV = koff.RPV/kon.RPV.
b Values obtained by globally fitting the data using KinTek Explorer.