TABLE 4.
Binding affinities of wild-type and mutant PDK2 proteins for lipL2 measured by ITC in the absence or presence of different ligands
The 500 μm concentration of lipL2 in the syringe was injected into the reaction cell containing 20 μm SUMO-PDK2 (based on the monomer) at 15 °C in the absence or presence of the indicated ligands (DCA, ADP, and/or ATP). The concentrations of these ligands are DCA at 0.3 mm and both ADP and ATP at 0.1 mm. All Kd values are expressed as average ± S.D. (n = 3). NM, not measurable.
| PDK2 | DCA (0.3 mm) | ADP (0.1 mm) | ATP (0.1 mm) | Kd | ΔH |
|---|---|---|---|---|---|
| μm | kcal/mol | ||||
| WT | − | − | − | 14.4 ± 2.7 | −16.1 ± 2.0 |
| WT | + | − | − | 14.3 ± 1.8 | −12.8 ± 2.2 |
| WT | − | + | − | 16.3 ± 3.5 | −4.5 ± 0.5 |
| WT | − | − | + | 16.1 ± 1.5 | −2.9 ± 0.5 |
| WT | + | + | − | NM | NM |
| WT | + | − | + | NM | NM |
| Y145F | − | − | − | 5.2 ± 0.1 | −1.5 ± 0.3 |
| Y145F | + | − | − | 5.2 ± 0.4 | −1.4 ± 0.4 |
| Y145F | − | + | − | NM | NM |
| Y145F | − | − | + | NM | NM |
| R149A | − | − | − | NM | NM |
| Y145F/R149A | − | − | − | NM | NM |
| D382A/W383A | − | − | − | NM | NM |