TABLE 1.
Steady-state and presteady-state kinetic parameters
| Steady state |
Titration, Kd,NADP+ | Presteady state |
||||||
|---|---|---|---|---|---|---|---|---|
| Km,TMA | Km,NADPH | kcat | kuncoupling | Kd,NADPH | kred | kox | ||
| μm | μm | s−1 | s−1 | μm | μm | s−1 | m−1s−1 | |
| WT | 7 | 18.6 | 3.3 | 0.5 | 21 | <10 | 18.4 | 65,000 |
| WTa | NDb | NDb | 0.4 | 0.2 | – | – | – | – |
| WTc | – | – | – | 0.13 | – | – | – | – |
| N78K | – | >160 | – | >0.07 | >250 | >250 | >0.1 | 670 |
| N78S | – | 3.6 | – | 0.7 | 22 | <10 | 6.7 | 5,000 |
| N78D | – | 85 | – | 0.19 | >250 | 270 | 1.9 | 2,200 |
a Performed using 150 μm thioNADPH and monitoring oxygen consumption.
b ND, not determined.
c Performed using 150 μm APADPH and monitoring oxygen consumption.