2′-O Methylation of Internal Adenosine by Flavivirus NS5 Methyltransferase
Figure 2
Optimal conditions for internal methylation for DENV-4 NS5 MTase.
SPA-based methylation assays were performed using uncapped pppA-RNA substrate (representing the first 211 nt of DENV genome). The reaction mixtures were incubated for 1 h at room temperature. Optimal pH, temperature, NaCl concentration, MgCl2 concentration, and MnCl2 concentration were obtained by titrating individual parameter while keeping other parameters at the optimal levels. Average results and standard deviations were obtained from three independent experiments.