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Steady-State Kinetic Parameters for MTHFS Mutants. All reactions were carried out in 100mM MES, pH 6.0 at 37°C.
| [6S] 5-formylTHF | [6R,S]-5-formylTHHF | ||||
|---|---|---|---|---|---|
| Km (μM) | kcat (min−1) | Km (μM) | kcat (min−1) | Ki (μM) | |
| WT-MTHFS | 10 ± 3 | 200 ± 50 | nd | nd | 0.7 ± 0.3 |
| Y151A | 9 ± 2 | 55 | 31 ± 2 | 0.2 | 0.3 ± 0.1 |
| Y152A | 3000 ± 500 | 1.2 ± 0.4 | nd | nd | -- |
| Y152F | 47 ± 15 | 258 ± 50 | -- | -- | -- |
nd – no activity detected
