Table 2. Binding Affinities and Thermodynamic Parameters of NanKs Binding to Their Ligands ManNAc and AMPPNP (ATP Analogue)b.
| NanK | in the presence of | ligand | Kd (μM) | ΔH (Kcal/mol) | TΔS (Kcal/mol) |
|---|---|---|---|---|---|
| P. multocida | ManNAc | 84 ± 9.9 | –1.763 ± 0.158 | 3.78 | |
| AMPPNP | no measurable binding | ||||
| AMPPNPa | ManNAc | 80 ± 1.9 | –13 ± 0.373 | –7.47 | |
| ManNAc | AMPPNP | 112 ± 14.5 | –4.3 ± 0.35 | 1.1 | |
| V. cholera | ManNAc | 39 ± 4.5 | –7.18 ± 0.825 | –1.174 | |
| AMPPNP | no measurable binding | ||||
| AMPPNPa | ManNAc | 27 ± 3.1 | –17 ± 0.9 | –10.787 | |
| ManNAc | AMPPNP | 44 ± 9.2 | –2.1 ± 0.482 | 3.814 | |
| H. influenzae | ManNAc | 33 ± 4.29 | –3.15 ± 0.2 | 2.947 | |
| AMPPNP | 53 ± 6.4 | –7.7 ± 2.44 | –1.85 | ||
| AMPPNPa | ManNAc | 28 ± 4.76 | –3.49 ± 0.475 | 2.717 | |
| ManNAc | AMPPNP | 72 ± 12 | –3.7 ± 0.68 | 1.94 | |
| F. nucleatum | ManNAc | 76 ± 14 | –0.814 ± 0.08 | 4.8 | |
| AMPPNP | 61 ± 7.48 | –3.613 ± 0.258 | 2.13 | ||
| AMPPNP | ManNAc | 55 ± 5.5 | –5.25 ± 0.51 | 0.548 | |
| ManNAc | AMPPNP | 35 ± 1.12 | –8.94 ± 0.18 | –2.87 | |
a
While most experiments were repeated multiple times, these titrations were only done once due to a paucity of AMPPNP.
b
Isothermal calorimetry (ITC) was used to measure the binding affinities, enthalpy, and entropy for the binding of NanKs with their ligands. The data were analyzed using Origin Analysis software.