Table 1.
The apparent Kd describing binding of wild-type RREIIB and RREII and their mutants to Rev-Fl obtained by fitting binding curves to a one-site binding model
| Apparent Kd (nM) | |
|---|---|
| RREIIB | |
| WT | 45.9 ± 15.2 |
| A68C | >2000 |
| U72C | >4000 |
| A68C/G50A/C69U | 33.0 ± 11.3 |
| m3U72 | 36.2 ± 9.3 |
| RREII | |
| WT | 1.5 ± 0.4 |
| A68C | >25 |
| U72C | >22 |
| A68C/G50A/C69U | 15.4 ± 5.1 |
Uncertainty represents standard deviation over triplicate measurements. The concentration of Rev-Fl peptide is 10 and 1 nM for RREIIB and RREII, respectively. Buffer conditions: 30 mM HEPES, pH = 7.0, 100 mM KCl, 10 mM sodium phosphate, 10 mM ammonium acetate, 10 mM guanidinium chloride, 2 mM MgCl2, 20 mM NaCl, 0.5 mM EDTA and 0.001% (v/v) Triton-X100