Figure 5
DEC formation by HIV-1 RT, TFV-terminated DNA 26/50-mer. (A) DEC formation by dCTP, FTC-TP, or EFV analyzed on a 6% non-denaturing polyacrylamide gel. (B) Quantification of the DEC formed by TFV-terminated DNA with HIV-1 RT in the presence of dCTP, FTC-TP, or EFV. Please note that the EFV x-axis is scaled significantly different from dCTP and FTC-TP. The amounts of free primer/template and DEC where quantified, and the percentage of DEC formed was plotted as a function of the concentration of dCTP, FTC-TP, or EFV. The solid line represents curve fitting of the data using the single ligand binding equation y = (Bmax × [ligand])/(Kd + [ligand]). The DEC formation by TFV-terminated DNA/RT in the presence of dCTP yielded Bmax = 49.2 ± 2.7% and Kd = 185 ± 28 μM (left panel); The DEC formation by TFV-terminated DNA/RT in the presence of FTC-TP with Bmax = 50.4 ± 1.1% and Kd = 52.6 ± 3.4 μM (middle panel); The DEC formation by TFV-terminated DNA/RT in the presence of EFV with Bmax = 48.0 ± 2.7% and Kd = 1.43 ± 0.25 μM (right panel).