Table S2.
kcatb,c (min−1) | KmRF b,c (μM) | Kib,c (μM) | kcat/KmRF b (min−1 μM−1) | kcatc,d (min−1) | KmATP c,d (μM) | kcat/KmATP d (min−1 μM−1) | |
---|---|---|---|---|---|---|---|
WT a | 301 | 13 | 4.0 | 23 | 68 | 14 | 4.9 |
H28A | 427 | 23 | 1.8 | 18 | 45 | 14 | 3.1 |
H28D | 287 | 25 | 1.9 | 12 | 55 | 12 | 4.7 |
H31A | 169 | 4.5 | 6.3 | 37 | 58 | 14 | 4.3 |
N125A | 111 | 1.7 | 4.6 | 65 | 46 | 24 | 1.9 |
N125D | 415 | 16 | 3.3 | 26 | 80 | 35 | 2.3 |
R161A | 300 | 12 | 5.1 | 25 | 65 | 12 | 5.6 |
R161D | 300 | 13 | 5.1 | 23 | 68 | 11 | 6.1 |
S164A | 259 | 8.8 | 4.4 | 29 | 59 | 12 | 5.0 |
S164D | 185 | 5.6 | 5.8 | 33 | 55 | 10 | 5.3 |
T165A | 171 | 10 | 3.2 | 17 | 66 | 12 | 5.4 |
T165D | 180 | 7.7 | 4.1 | 23 | 45 | 11 | 4.1 |
Data from [24].
Parameters determined at saturated ATP concentrations.
Inhibition by substrate prevented the determination of true parameters and the values here reported correspond to apparent constants; appkcat and appKm. Estimated errors in appkcat and appKm values increased up to ±35%.
Parameters estimated using a RF concentration exhibiting ~80% maximal activity before the maximum experimentally detected. Errors in kcat and Km were considered within ±10%, being this value taken larger than the standard deviation between three independent experiments and the numerical error after fitting analysis of each experiment to the Michaelis-Menten equation.