Table 1.
Binding affinities and kinetic parameters of the ligands for hPGK in the absence and presence of counter ligands*
| Ligand [Ex-model]† |
KD/mM | α | ΔΔG/kJ mol−1 | koff/s−1 | ||
| −3PG | +3PG | −3PG | +3PG | |||
| MgADP [F to I] |
0.224 (0.216 to 0.232) |
0.501 (0.477 to 0.527) |
0.448 (0.426 to 0.470) |
2.07 (1.94 to 2.20) |
811 (749 to 880) |
1,348 (1,227 to 1,493) |
|
0.237
(0.203 to 0.266) |
0.320
(0.297 to 0.354) |
0.740
(0.671 to 0.797) |
0.775
(0.584 to 1.03) |
— | — | |
| MgAMPPNP [F] |
0.894 (0.852 to 0.942) |
0.656 (0.624 to 0.687) |
1.36 (1.30 to 1.43) |
−0.791 (−0.920 to −0.675) |
N/A‡ | N/A‡ |
|
1.08
(1.04 to 1.17) |
0.737
(0.663 to 0.806) |
1.47
(1.34 to 1.63) |
−0.990 (−1.26 to −0.759) |
— | — | |
| −nucleotide | +MgADP | α | ΔΔG/kJ mol−1 | |||
| 3PG [F] |
0.112 (0.106 to 0.119) 0.158 (0.140 to 0.175) |
0.251 (0.239 to 0.263) |
0.448 (0.426 to 0.470) |
2.07 (1.94 to 2.20) |
N/A‡ | N/A‡ |
|
0.213
(0.198 to 0.235) |
0.740
(0.671 to 0.797) |
0.775
(0.584 to 1.03) |
— | — | ||
| +MgAMPPNP | α | ΔΔG/kJ mol−1 | ||||
| 0.0823 (0.0782 to 0.0862) |
1.36 (1.30 to 1.43) |
−0.791 (−0.920 to −0.675) |
N/A‡ | N/A‡ | ||
|
0.108
(0.0967 to 0.118) |
1.47
(1.34 to 1.63) |
−0.990 (−1.26 to −0.759) |
— | — | ||
*The values obtained from the NMR data are presented in the upper rows, whereas those obtained from the ITC data are displayed in the lower rows with italics for each ligand. The KD values in the presence of counter ligands are calculated from these α-values (Fig. 1C). Values in brackets represent the 95% bootstrap CIs.
†Exchange model (Ex-model) used for the global CS-fitting analyses are shown in square brackets. [F to I] and [F] indicate “fast-to-intermediate” and “fast” exchanges, respectively.
‡Not applicable owing to fast exchange.