TABLE 5.
Apparent dissociation constants calculated for PTB1 and PTB1:12: HCV X RNAs
| RNA | ||
| Protein | X RNA | X SLIII |
| PTB1 |
KD1 = 800 nM, n = 1 KD2 = 1 μM, n = 1 |
KD = 285 nM ± 120 nM |
| PTB1:12 |
KD1 = 800 nM, n = 1 KD2 = 1 μM, n = 1 |
KD = 1.6 μM ± 0.6 μM |
For protein binding to X RNA, dissociation constants were calculated from the relation P = 1 + (K1[P])n1 + (K1K2 [P]2)n2, where n is the Hill coefficient, using Origin 7.5 (OriginLab, Northampton, MA). Binding to X SLIII was measured using nitrocellulose filter binding, and the data were fit by Kaleidegraph to KD = ([R] – [PR])([P] – [PR])/[PR]; 100 mM NaCl, 1 mM MgCl2 , 10 mM cacodylate (pH 7), 20 μg/mL BSA, 10 μg/mL tRNA.