Table 2. Summary of thermodynamic-binding parameters for complex formation between different H3 peptides and WT and mutant BAZ2A/B PHD fingers.
Error values reported on dissociation constant (KD), stoichiometry of binding (N) and binding enthalpy (ΔH) are generated by the Origin program and reflect the quality of the fit between the nonlinear least-squares curve and the experimental data. Errors reported on TΔS and ΔG were propagated from the errors of KD and ΔH. Raw ITC data are shown for each titration in Supplementary Data (Supplementary Figure S1).
| Peptide | Protein | KD (µM) | N | ΔH (kcal/mol) | TΔS (kcal/mol) | ΔG (kcal/mol) |
|---|---|---|---|---|---|---|
| ARTK | BAZ2A PHD | 100 ± 4 | 1.57 ± 0.03 | −6.1 ± 0.2 | −0.6 ± 0.2 | −5.46 ± 0.02 |
| ARTA | BAZ2A PHD | 56 ± 2 | 1.44 ± 0.01 | −4.7 ± 0.1 | 1.1 ± 0.1 | −5.81 ± 0.02 |
| ARTKQ | BAZ2A PHD | 210 ± 30 | 1.6 ± 0.1 | −3.7 ± 0.4 | 1.3 ± 0.4 | −5.0 ± 0.1 |
| ARTKQTARKS | BAZ2A PHD | 51 ± 2 | 1.35 ± 0.01 | −5.8 ± 0.1 | 0.1 ± 0.1 | −5.86 ± 0.02 |
| AATKQTARKS | BAZ2A PHD | >1000 | N.D. | N.D. | N.D. | N.D. |
| ARAKQTARKS | BAZ2A PHD | No binding | ||||
| ARTAQTARKS | BAZ2A PHD | 42 ± 2 | 1.03 ± 0.02 | −6.1 ± 0.2 | −0.1 ± 0.2 | −6.00 ± 0.03 |
| ARTKATARKS | BAZ2A PHD | 9.3 ± 0.2 | 1.21 ± 0.01 | −6.13 ± 0.02 | 0.74 ± 0.02 | −6.87 ± 0.01 |
| ARTKQAARKS | BAZ2A PHD | 55 ± 2 | 0.90 ± 0.01 | −5.6 ± 0.1 | 0.2 ± 0.1 | −5.82 ± 0.02 |
| ARTAATARKS | BAZ2A PHD | 12.1 ± 0.3 | 1.32 ± 0.01 | −7.01 ± 0.04 | −0.30 ± 0.04 | −6.71 ± 0.02 |
| ARTGGTARKS | BAZ2A PHD | 143 ± 7 | 1.28 ± 0.04 | −7.4 ± 0.3 | −2.2 ± 0.3 | −5.25 ± 0.03 |
| ARTKQ | BAZ2B PHD | 190 ± 25 | 1.0 ± 0.1 | −4.7 ± 0.8 | 0.4 ± 0.8 | −5.1 ± 0.1 |
| ARTKQTARKS | BAZ2B PHD | 40.0 ± 0.5 | 1.45 ± 0.04 | −5.43 ± 0.02 | 0.58 ± 0.02 | −6.01 ± 0.01 |
| AATKQTARKS | BAZ2B PHD | >1000 | N.D. | N.D. | N.D. | N.D. |
| ARAKQTARKS | BAZ2B PHD | >5000 | N.D. | N.D. | N.D. | N.D. |
| ARTAQTARKS | BAZ2B PHD | 8.5 ± 0.2 | 0.88 ± 0.01 | −6.62 ± 0.04 | 0.30 ± 0.05 | −6.92 ± 0.01 |
| ARTKATARKS | BAZ2B PHD | 7.0 ± 0.2 | 1.20 ± 0.03 | −6.18 ± 0.02 | 0.86 ± 0.03 | −7.04 ± 0.02 |
| ARTKQAARKS | BAZ2B PHD | 44.6 ± 0.8 | 1.01 ± 0.01 | −5.3 ± 0.1 | 0.68 ± 0.06 | −5.94 ± 0.01 |
| ARTAATARKS | BAZ2B PHD | 2.6 ± 0.1 | 1.13 ± 0.01 | −7.12 ± 0.02 | 0.50 ± 0.03 | −7.63 ± 0.02 |
| ARTGGTARKS | BAZ2B PHD | 166 ± 8 | 1.3 ± 0.1 | −8.4 ± 0.5 | −3.3 ± 0.5 | −5.16 ± 0.03 |
Abbreviations: N.D.: not determined.