Table 1.
UbaA Variant | Ligand | Tm (°C) |
Tm Shift (°C) |
---|---|---|---|
wt | -a | 58.60 ± 1.13 | - |
ATP | 66.80 ± 3.96 | 8.20 ± 4.12 | |
AMP | 59.90 ± 0.42 | 1.30 ± 1.21 | |
ADP | 63.00 ± 0.85 | 4.40 ± 1.41 | |
AMP-PNP | 66.00 ± 3.39 | 7.40 ± 3.58 | |
CTP | 57.60 ± 1.13 | -1.00 ± −1.60 | |
GTP | 59.60 ± 1.41 | 1.00 ± 1.81 | |
TTP | 59.70 ± 2.12 | 1.10 ± 2.40 | |
UTP | 59.70 ± 0.99 | 1.10 ± 1.50 | |
| |||
K87R | - | 59.68 ± 1.42 | - |
ATP | 60.12 ± 0.46 | 0.44 ± 1.61 | |
| |||
D131N | - | 61.12 ± 1.85 | - |
ATP | 60.92 ± 3.02 | −0.20 ± 3.54 | |
| |||
C188A | - | 56.08 ± 2.04 | - |
ATP | 63.20 ± 1.11 | 7.12 ± 2.32 |
-, no ligand control. Means of thermal shift difference and standard deviations were calculated (n = 2 for the nucleotide ligands, n = 6 for the UbaA variants). See Methods for assay details.