Figure 4
Determination of binding specificity of fragment 204 to NS5B in solution.
(A) LC-MS chromatograms of fragment 204 detected from incubations containing the wild-type or specific mutants of NS5B, or from the protein-free control experiment. M414 and C366 are two residues observed to interact with the fragment in the crystal structure. M423 is speculated to have no interaction with the fragment. (B) Relative quantification for fragment 204 based on its MS intensity from (A) implies its interactions with M414T and C366A were significantly impaired whereas interaction with M423T was not, an evidence of binding specificity at these two sites. MS intensity percentages of fragment 204 from the mutant incubation relative to the wild-type incubation (defined as 100%) were averages from duplicate measurement and s.d. are shown accordingly.
