Table 1.
Summary of thermodynamic and curve fitting parameters for ITC assays using recombinant PHF1 or PHF19 Tudor domain proteins and various histone peptides. See also Figures S1L-S and S2G.
| Protein | Peptide | ΔH(cal/mol) | ΔS (cal/mol/deg) |
Ka(M-1) | N | Chi2/ DoF |
Kd (µM) |
|---|---|---|---|---|---|---|---|
| PHF1Tudor, wt | H3(28–43)K36me3 | −5626 ± 108 | 4.45 | 2.38E5 ± 1.91E4 | 0.79 ± 0.01 | 5415 | 4.2 ± 0.3 |
| PHF1Tudor, wt | H3(28–43)K36me2 | −3994 ± 261 | 7.26 | 5.18E4 ± 6.79E3 | 0.94 ± 0.04 | 4338 | 19.3 ± 2.5 |
| PHF1Tudor, wt | H3(28–43)K36me1 | −817 ± 342 | 17.3 | 2.63E4 ± 1.12E4 | 0.70 ± 0.23 | 658.3 | 38.0 ± 16.1 |
| PHF1Tudor, wt | H3(28–43)K36me0 | -- | -- | -- | -- | -- | N.D |
| PHF1Tudor, wt | H3(19–35)K27me3 | −5141 ± 469 | −1.21 | 5.93E3 ± 486 | 1.03 ± 0.07 | 347.7 | 168 ± 13.8 |
| PHF1Tudor, wt | H3(1–15)K9me3 | −817 ± 343 | 17.3 | 2.63E4 ± 1.12E4 | 0.70 ± 0.23 | 658.3 | 38.0 ± 16.2 |
| PHF1Tudor, W41A | H3(28–43)K36me3 | −1355 ± 457 | 15.8 | 3.32E4 ± 1.20E4 | 0.68 ± 0.18 | 1456 | 30.1 ± 10.9 |
| PHF1Tudor, Y47A | H3(28–43)K36me3 | -- | -- | -- | -- | -- | N.D. |
| PHF1Tudor, F65A | H3(28–43)K36me3 | -- | -- | -- | -- | -- | N.D. |
| PHF1Tudor, F71A | H3(28–43)K36me3 | -- | -- | -- | -- | -- | N.D. |
| PHF1Tudor, E66K | H3(28–43)K36me3 | −6471 ± 229 | −1.94 | 4.39E4 ± 3.76E3 | 0.60 ± 0.02 | 5055 | 22.8 ± 2.0 |
| PHF19Tudor, wt | H3(27–46)K36me3 | −6610 ± 267 | 0.466 | 1.60E5 ± 1.65E4 | 0.58 ± 0.02 | 7436 | 6.2 ± 0.6 |
| PHF19Tudor, wt | H3(27–46)K36me1 | -- | -- | -- | -- | -- | N.D. |
| PHF19Tudor, Y56A | H3(27–46)K36me3 | -- | -- | -- | -- | -- | N.D. |
wt, wildtype; N.D, not determined.