Table 1.
Binding affinities of mono-, di-, tri-, and oligonucleotide probes for eIF4E together with fluorescence enhancement factors (g).
| L.p | KD / nM | g = FLbound / FLfree |
|---|---|---|
| 1a | 59.7 ± 0.9 | 0.90 ± 0.02 |
| 1b | 10.5 ± 1.0 | 0.98 ± 0.01 |
| 1c | 20.7 ± 5.9 | 0.77 ± 0.02 |
| 1e | 41.3 ± 4.4 | 1.10 ± 0.03 |
| 1f | 63.7 ± 15.3 | 1.17 ± 0.07 |
| 2a | 128.1 ± 11.0 | 1.10 ± 0.06 |
| 2b | 108.6 ± 6.2 | 0.96 ± 0.04 |
| 2c | 106.4 ± 7.2 | 0.91 ± 0.02 |
| 2d | 79.8 ± 3.5 | 0.99 ± 0.05 |
| 3a | 68.3 ± 14.4 | 0.56 ± 0.01 |
| 3b | 33.9 ± 6.3 | 1.44 ± 0.04 |
| 3c | 45.7 ± 7.4 | 1.15 ± 0.02 |
| 4a | 477.4 ± 29.8 | 0.93 ± 0.02 |
FA experiments were performed in black 96-well plates using a Biotek Synergy H1 plate reader. Each well (200 µL) contained a fluorophore-tagged probe (0.5, 1, 2, or 10 nM) and increasing concentrations of the desired protein (from 0 to 2.5 µM).