Table 3.
Time-resolved anisotropy of free and bound PTB/W proteinsa
| Protein | βf | Φf (ns) | βs | Φs (ns) | Χ2 | <τ> (ns) |
|---|---|---|---|---|---|---|
| PTB1/W1 | 0.62 | 0.11 ± 0.08 | 0.38 | 7.3 ± 1.2 | 1.2 | 4.4 |
| + GABA RNA11@ |
0.31 | 0.10 ± 0.07 | 0.69 | 7.2 ± 0.52 | 1.2 | 4.0 |
| + HCV X RNA | 0.72 | 0.12 ± 0.07 | 0.28 | 7.9 ± 1.6 | 1.1 | 5.0 |
| PTB1/W3 | 0.53 | 0.23 ± 0.12 | 0.46 | 5.4 ± 1.2 | 1 | 3.9 |
| + GABA RNA11 |
0.67 | 0.25 ± 0.21 | 0.33 | 4.7 ± 1.5 | 1.08 | 4.3 |
| + HCV X RNA | 0.87 | 0.33 ± 0.11 | 0.13 | 9.1 ± 1.7 | 1.04 | 4.3 |
| + HCV X SL3 | 0.90 | 0.14 ± 0.04 | 0.10 | 21.6 ± 5.9 | 1.04 | 3.8 |
| PTB1/W4 | 0.63 | 0.38 ± 0.19 | 0.37 | 8.4± 2.2 | 1.07 | 5.6 |
| + GABA RNA11 |
0.62 | 0.34 ± .23 | 0.38 | 9.8 ± 3.3 | 1.02 | 5.3 |
| + HCV X RNA | 0.71 | 0.5± 0.2 | 0.29 | 17.5 ± 3.8 | 1.2 | 5.4 |
| + HCV X SL 3 | 0.73 | 0.23 ± 0.16 | 0.27 | 9.4 ± 3.2 | 1.1 | 5.5 |
a
Protein alone concentrations range from 1 to 5 µM in 100 mM NaCl, 10 mM sodium cacodylate pH 7.5, 10 mM DTT, room temperature.
@
250 mM NaCl only. RNA concentrations were 1 µM, and protein concentrations were adjusted to saturate the binding ([P]/[R] = 1 for HCV X SL3; [P]/[R] = 2 or HCV X and GABA RNA 11). Data acquired on a time-correlated single photon counting instrument, and analyzed with PicoQuant FluoFit. Steady state fluorescence spectra were collected before and after time-resolved experiments. <τ> values from Supplementary Table.