Table 1.
Equilibrium dissociation constants for binding of CST and telomerase to 3xTEL DNA by EMSA at 4°C and by FP at room temperature. Each value is the mean ± SD of n independent experiments. Relative Kd values are normalized to WT CST. For the weak-binding CST mutants, the FP binding curves did not go to completion, so the Kd values are approximate.
| EMSA | FP | ||||||
|---|---|---|---|---|---|---|---|
| Binders | Mutations | K d,app. (nM) | Relative Kd | n | K d,app. (nM) | Relative Kd | n |
| WT CST | NA | 6.5 ± 4.4 | 1.0 | 9 | 2.2 ± 0.28 | 1.0 | 4 |
| G1.1 CST | Y949A, E951K | 133 ± 73 | 20 | 6 | 520 ± 80 | 230 | 2 |
| G2.1 CST | R978E, N981D, Y983A | 210 ± 106 | 32 | 5 | 800 ± 220 | 360 | 2 |
| G3.1 CST | K1164E, K1167E | 97 ± 27 | 15 | 3 | 420 ± 290 | 190 | 2 |
| G4.1 CST | K743E, R744E | 7.7 ± 3.4 | 1.1 | 3 | 2.0 ± 0.16 | 0.90 | 2 |
| Telomerase | NA | – | – | – | 0.54 ± 0.25 | 0.25 | 4 |