Table 1.
Steady-state and pre-steady-state kinetic parameters of WT hAIF∆1-101 and its W196 variants.
| hAIF | Steady-state | Pre-steady-sate | |||||||
|---|---|---|---|---|---|---|---|---|---|
|
k
cat
(s−1) |
K
m
NADH
(μM) |
k
cat/KmNADH (s−1mM−1) |
k
HT
(s−1) |
K
d
NADH
(μM) |
k
HT/KdNADH (s−1mM−1) |
k
CTC
1
(s−1) |
K
d
NAD+
(μM) |
k
CTC/KdNAD+ (s−1mM−1) |
|
| WT | 0.9 ± 0.1 | 495 ± 170 | 1.9 ± 0.9 | 1.5 ± 0.1 | 4090 ± 300 | 0.4 ± 0.1 | 45 ± 2 | 2080 ± 250 | 22 ± 3 |
| W196Y | 2.7 ± 0.1 | 505 ± 35 | 5.3 ± 0.6 | 12 ± 1 | 2870 ± 320 | 4.0 ± 0.8 | 16 ± 1 | 183 ± 27 | 87 ± 14 |
| W196L | 2.8 ± 0.1 | 187 ± 18 | 15 ± 1.9 | 36 ± 1 | 1725 ± 210 | 21 ± 3 | 29 ± 2 | 433 ± 11 | 67 ± 18 |
| W196A | 4.3 ± 0.2 | 25 ± 4 | 172 ± 35 | 126 ± 1 | 1070 ± 40 | 117 ± 5.3 | 30 ± 1 | 394 ± 50 | 76 ± 10 |
Assays were performed at 25°C in 50 mM potassium phosphate, pH 7.4 (n = 3, mean ± SD). 1Kinetic parameters for CTC formations were obtained with hAIF∆1-101phrd variants.