Table 1.
Kinetic Parameters of the Wild-Type and Mutant Maf Proteins
| Protein | Variable substrate | KM (μM) | kcat (s−1) | kcat/KM (s−1M−1) |
|---|---|---|---|---|
| YhdE | ||||
| WT | dTTP | 53.0 ± 2.0 | 15.4 ± 0.3 | 2.9 × 105 |
| WT | UTP | 69.1 ± 3.1 | 15.2 ± 0.5 | 2.2 × 105 |
| WT | CTP | 105.9 ± 10.8 | 4.9 ± 0.2 | 4.6 × 104 |
| WT | m5UTP | 32.0 ± 2.2 | 18.2 ± 0.7 | 5.7 × 105 |
| WT | m5CTP | 44.8 ± 2.2 | 9.2 ± 0.2 | 2.1 × 105 |
| WT | pseudoUTP | 47.2 ± 1.5 | 10.5 ± 0.2 | 2.2 × 105 |
| S10A | dTTP | 78.9 ± 3.5 | 4.5 ± 0.1 | 5.7 × 104 |
| R12A | dTTP | 40.9 ± 2.0 | 5.5 ± 0.1 | 1.3 × 105 |
| K82A | dTTP | 36.1 ± 2.0 | 0.8 ± 0.1 | 2.2 × 104 |
| BSU28050 | ||||
| WT | dTTP | 50.6 ± 2.1 | 11.6 ± 0.2 | 2.3 × 105 |
| WT | UTP | 72.4 ± 5.2 | 4.9 ± 0.2 | 6.8 × 104 |
| WT | CTP | 25.0 ± 0.8 | 7.1 ± 0.1 | 2.8 × 105 |
| WT | m5UTP | 4.7 ± 0.2 | 2.4 ± 0.1 | 5.1 × 105 |
| WT | m5CTP | 7.8 ± 0.2 | 8.5 ± 0.1 | 1.1 × 106 |
| WT | pseudoUTP | 5.7 ± 0.3 | 7.2 ± 0.2 | 1.3 × 106 |
| YOR111W | ||||
| WT | dTTP | 47.4 ± 2.3 | 5.7 ± 0.1 | 1.2 × 105 |
| WT | UTP | 29.7 ± 1.4 | 2.6 ± 0.1 | 8.8 × 104 |
| WT | CTP | 72.0 ± 2.6 | 1.2 ± 0.02 | 1.7 × 104 |
| WT | m5UTP | 13.0 ± 0.3 | 2.2 ± 0.1 | 1.7 × 105 |
| WT | pseudoUTP | 20 ± 0.4 | 5.3 ± 0.1 | 2.7 × 105 |
| T26A | dTTP | 72.5 ± 5.2 | 4.1 ± 0.1 | 5.7 × 104 |
| R29A | dTTP | 39.4 ± 0.8 | 6.7 ± 0.1 | 1.7 × 105 |
| Q76A | dTTP | 60.7 ± 4.7 | 3.7 ± 0.1 | 6.1 × 104 |
| N77A | dTTP | 56.9 ± 3.7 | 3.1 ± 0.1 | 5.4 × 104 |
| E188A | dTTP | 42.9 ± 2.1 | 1.4 ± 0.1 | 3.3 × 104 |
| F193A | dTTP | 62.3 ± 2.0 | 3.4 ± 0.1 | 5.5 × 104 |
| K194A | dTTP | 52.1 ± 3.5 | 4.1 ± 0.1 | 7.9 × 104 |
| ASMTL-Maf | ||||
| WT | dTTP | 32.7 ± 3.4 | 0.7 ± 0.1 | 2.1 × 104 |
| WT | UTP | 41.2 ± 9.3 | 0.7 ± 0.1 | 1.7 × 104 |
| WT | CTP | 17.4 ± 0.8 | 0.8 ± 0.02 | 4.6 × 104 |
| WT | dCTP | 22.8 ± 1.6 | 0.3 ± 0.01 | 1.3 × 104 |
| WT | m5UTP | 16.1 ± 1.5 | 1.5 ± 0.1 | 9.3 × 104 |
| WT | m5CTP | 39.4 ± 6.5 | 3.7 ± 0.3 | 9.4 × 104 |
| WT | pseudoUTP | 18.7 ± 3.8 | 2.5 ± 0.2 | 1.3 × 105 |
| WT | 8-oxo-GTP | 10.7 ± 0.6 | 0.2 ± 0.01 | 1.9 × 104 |
| WT | N4-m-dCTP | 10.3 ± 0.7 | 0.3 ± 0.01 | 2.9 × 104 |
| R23A | dTTP | 27.7 ± 1.0 | 1.6 ± 0.1 | 5.8 × 104 |
| YceF | ||||
| WT | m7GTP | 32.8 ± 3.1 | 1.2 ± 0.1 | 3.7 × 104 |
| W12A | m7GTP | 217.2 ± 42.3 | 6.4 ± 1.1 | 2.9 × 104 |
| Q70A | m7GTP | 173.3 ± 11.5 | 5.0 ± 0.3 | 2.9 × 104 |
| E154A | m7GTP | 79.0 ± 17.4 | 2.1 ± 0.4 | 2.7 × 104 |
| STM1189 | ||||
| WT | m7GTP | 30.9 ± 1.2 | 1.5 ± 0.1 | 4.9 × 104 |
The assays were performed in the presence of saturating concentrations of Co2+ (2 mM) at 30°C (for BSU28050 and YOR111W) or 37°C (for other proteins). WT, wild type. See also Figure S4.