Table 1. Kinetic constants of UNG2 and N-terminal UNG2 phosphomimicking mutants.
Enzyme | Substrate | Km (μM) | kcat (min−1) |
---|---|---|---|
Without RPA | |||
WT | U:A | 1.0±0.08 | 449±9 |
S23D | U:A | 1.8±0.06 | 870±6 |
S23D, T60D | U:A | 1.4±0.06 | 641±6 |
S23D, T60D, S64D | U:A | 1.2±0.07 | 726±5 |
WT | Uss | 0.05±0.00 | 663±10 |
S23D | Uss | 0.14±0.01 | 1981±13 |
S23D, T60D | Uss | 0.09±0.01 | 1141±15 |
S23D, T60D, S64D | Uss | 0.11±0.01 | 1360±10 |
With RPA | |||
WT | U:A | 0.59±0.09 | 143±5 |
S23D | U:A | 0.70±0.06 | 187±7 |
S23D, T60D | U:A | 0.66±0.06 | 162±7 |
S23D, T60D, S64D | U:A | 0.67±0.07 | 272±6 |
WT | Uss | 0.04±0.005 | 773±18 |
S23D | Uss | 0.05±0.004 | 1204±11 |
S23D, T60D | Uss | 0.03±0.004 | 546±11 |
S23D, T60D, S64D | Uss | 0.02±0.003 | 882±10 |