Table 4.
Calorimetric binding data for native L3MBTL3 and RBPJ mutants
| RBPJ | K (M−1) | K d (μM) | ΔG° (kcal/mol) | ΔH° (kcal/mol) | –TΔS° (kcal/mol) | ΔΔG° (kcal/mol) | |
|---|---|---|---|---|---|---|---|
| WT | 1.1 ± 0.1 × 106 | 0.92 | –8.3 ± 0.1 | –13.1 ± 0.3 | 4.8 ± 0.3 | – | |
| BTD mutants | E260A | 2.6 ± 0.8 × 105 | 3.8*** | –7.4 ± 0.2 | –13.6 ± 3.4 | 6.2 ± 3.6 | 0.9 |
| E260Kψ | 2.2 ± 0.1 × 105 | 4.5** | –7.3 ± 0.1 | –16.8 ± 0.9 | 9.5 ± 0.9 | 1.0 | |
| F261A | NBD | — | — | — | — | — | |
| V263A | 3.3 ± 0.9 × 105 | 3.0*** | –7.5 ± 0.2 | –10.3 ± 2.1 | 2.8 ± 2.3 | 0.8 | |
| K275M | 2.4 ± 0.6 × 105 | 4.2*** | –7.3 ± 0.1 | –13.7 ± 1.4 | 6.4 ± 1.5 | 1.0 | |
| A284V | NBD | — | — | — | — | — | |
| Q333A | 7.6 ± 1.5 × 105 | 1.3* | –8.0 ± 0.1 | –10.6 ± 1.0 | 2.6 ± 0.9 | 0.3 |
Constructs: L3MBTL3 (52–70) and RBPJ (53–474). All experiments were performed at 25°C. NBD represents no binding detected. ψN = 2 replicates. Values are the mean of at least three independent experiments and errors represent the standard deviation of multiple experiments. ΔΔG° = ΔG°(mutant) – ΔG°(mutant). P-value determined from a two-tailed t-test (***P < 0.001, **P < 0.01, *P < 0.05 and ns, not significant), comparing WT RBPJ to point mutants.