Table 2.
SpNatC complex (WT and mutant SpNaa30) catalytic parameters
| Substrate | Protein (SpNatC) | kcat* (min−1) | Relative kcat (normalized to WT) | Km* (μM) | Relative Km (normalized to WT) | kcat/Km (normalized to WT) |
|---|---|---|---|---|---|---|
|
Gag Peptide (MLRF-) |
WT | 19.7 ± 3.6 | 1.0 | 7.8 ± 1.4 | 1.0 | 1.0 |
| S24A | 19.1 ± 2.8 | 0.97 | 8.9 ± 1.0 | 1.1 | 0.85 | |
| E25A | 33.6 ± 11 | 1.7 | 11.1 ± 6.1 | 1.42 | 1.2 | |
| Y27A | 8.7 ± 1.6 | 0.44 | 2.5 ± 0.4 | 0.32 | 1.4 | |
| Y71A | 32.7 ± 8.5 | 1.7 | 10.7 ± 3.8 | 1.4 | 1.2 | |
| Y71F | 18.4 ± 8.3 | 0.94 | 13.9 ± 11 | 1.8 | 0.52 | |
| M74A | 13.3 ± 1.9 | 0.68 | 11.4 ± 5.2 | 1.5 | 0.46 | |
| E109A | 10.4 ± 0.6 | 0.53 | 15.2 ± 9.6 | 1.95 | 0.27 | |
| E109Q | 2.4 ± 0.4 | 0.12 | 72.1 ± 95 | 9.25 | 0.013 | |
| N114A | 10.9 ± 1.9 | 0.55 | 4.4 ± 0.9 | 0.56 | 0.99 | |
| Y121A | 1.8 ± 0.7 | 0.093 | 9.7 ± 5.0 | 1.2 | 0.075 | |
| Y121F | 1.8 ± 0.5 | 0.090 | 5.3 ± 2.0 | 0.68 | 0.13 | |
| Y135A | 4.1 ± 1.3 | 0.21 | 4.0 ± 3.9 | 0.51 | 0.41 | |
| Y136A | 0.2 ± 0.1 | 0.011 | 0.7 ± 0.4 | 0.091 | 0.13 |
*
values indicated mean ± S.D. of three independent kinetic curves fitting results.
See Supplementary Figure.6, for the raw curves.