Table 1. Analysis of Binding Interactions between Gankyrin and FtsH-S6 ATPase Proteins by ITCa.
| entry | gankyrin | FtsH-S6 ATPase | KD (nM) | ΔG (kcal/mol) | ΔH (kcal/mol) | –TΔS (kcal/mol) |
|---|---|---|---|---|---|---|
| 1 | wild-type | wild-type | 67.3 ± 5.7 | –9.8 ± 0.1 | –28.7 ± 0.5 | 19.0 ± 0.6 |
| 2 | wild-type | R342A | 216.6 ± 25.8 | –9.1 ± 0.1 | –22.0 ± 0.8 | 12.9 ± 0.8 |
| 3 | wild-type | R338A/R342A | 2549 ± 353 | –7.6 ± 0.1 | –6.1 ± 0.7 | –1.5 ± 0.8 |
| 4 | wild-type | R338A/R339A/R342A | 7471 ± 301 | –7.0 ± 0.1 | –2.2 ± 0.1 | –4.7 ± 0.1 |
| 5 | wild-type | E356A/E357A | 71.8 ± 5.9 | –9.8 ± 0.1 | –27.3 ± 0.9 | 17.5 ± 0.8 |
| 6 | wild-type | D359A/D362A | no binding | – | – | – |
| 7 | wild-type | K397E | 95.2 ± 12.2 | –9.7 ± 0.2 | –25.6 ± 2.5 | 15.9 ± 2.7 |
| 8 | R41A | wild-type | 313.3 ± 17.6 | –8.1 ± 1.2 | –17.1 ± 1.7 | 9.0 ± 2.8 |
| 9 | K116A | wild-type | 71.3 ± 15.5 | –9.7 ± 0.2 | –24.0 ± 0.9 | 14.3 ± 1.1 |
| 10 | D39A/D71A | wild-type | 93.0 ± 5.6 | –9.7 ± 0.2 | –25.0 ± 0.7 | 15.4 ± 0.8 |
| 11 | R41A/K116A | wild-type | 3633 ± 404 | –7.4 ± 0.1 | –4.9 ± 0.6 | –2.5 ± 0.7 |
| 12 | E182A | wild-type | 140.6 ± 9.7 | –9.4 ± 0.1 | –28.2 ± 2.1 | 18.8 ± 2.0 |
a
All errors represent the standard deviation of three separate experiments. ITC conditions were as follows: 20 mM sodium phosphate, 150 mM NaCl, and 2.5 mM 2-mercaptoethanol (pH 7.4) at 25 °C.