Table 3.
The summary of ITC thermodynamic parameters. The binding curves and plots are presented in Supplementary Figure S5 and Figure 6G and H. * the binding was performed in the presence of 100 μM synthetic t6A
| Titrant (μM) | Injectant (μM) | N (sites) | K D (μM) | ΔH (kcal/mol) | ΔG (kcal/mol) |
|---|---|---|---|---|---|
| TsaN1−392 (30) | ATPγS (360) | 1.18 ± 0.14 | 10.9 ± 8.91 | −6.02 ± 2.38 | −6.77 |
| ADP (360) | 1.10 ± 0.10 | 5.55 ± 3.53 | −2.74 ± 0.62 | −7.05 | |
| AMP (360) | 1.15 ± 0.14 | 14.5 ± 7.41 | −4.74 ± 1.46 | −6.49 | |
| A (360) | No binding | ||||
| TsaN(1−392)H155A/H159A (30) | ATP (360) | No binding | |||
| TsaN749−1008 (30) | ATP (360) | No binding | |||
| ADP (360) | No binding | ||||
| AMP (360) | No binding | ||||
| A (360) | No binding | ||||
| t6A (360) | 0.80 ± 0.03 | 9.14 ± 2.06 | −6.02 ± 0.62 | −6.76 | |
| AMPCPP (360)* | No binding | ||||
| TsaN(749−1008)R814A (30) | t6A (360) | 1.02 ± 0.06 | 10.6 ± 4.13 | −8.98 ± 1.49 | −6.67 |
| TsaN(749−1008)K827A (30) | t6A (360) | 0.94 ± 0.04 | 11.2 ± 3.26 | −9.38 ± 1.25 | −6.65 |
| EcTcdA (30) | t6A (360) | No binding | |||