Table 3. The Dissociation Constants (Kd, μM) for Impα:NLS Interactions.
| GST-NLS | mImpαΔIBB | yImpαΔIBB | rImpα1aΔIBB | mImpαΔIBB D192K | mImpαΔIBB E396R | rImpα1a | rImpα1aΔIBB D188K | rImpα1aΔIBB E388R |
|---|---|---|---|---|---|---|---|---|
| SV40TAg | 0.0070 ± 0.0019 | 0.0110 ± 0.0024 | 0.0040 ± 0.0002 | 3.91 ± 0.27 | 0.053 ± 0.009 | 1.73 ± 0.15 | 0.680 ± 0.094 | 0.0470 ± 0.008 |
| A89 | 2.90 ± 0.36 | 1.70 ± 0.20 | 0.140 ± 0.023 | Not detected | 7.65 ± 1.54 | 5.95 ± 1.63 | 0.410 ± 0.096 | 25.0 ± 8.5 |
| B54 | 1.70 ± 0.23 | 1.80 ± 0.20 | 0.025 ± 0.004 | 5.61 ± 2.00 | 4.95 ± 0.56 | 0.118 ± 0.017 | 0.0270 ± 0.0017 | 0.137 ± 0.019 |
The Kd values (in μM) were calculated using program GraphPad (Prism). Each assay was performed in triplicate, and the values with se correspond to the best fit to the one-site specific binding equation [Y = Bmax*X/(Kd + X), where Bmax is the maximum specific binding with the same unit as Y, Kd is the equilibrium binding constant, and X is ligand concentration]. The calculation assumed one-site binding to allow the comparison of overall binding affinities between different samples. The binding isotherms of representative measurements are provided in the Supplemental Figure 7 online.