Figure 6.
PepA binds both ssRNA and dsRNA. Filter binding assays were performed to test binding of PepA to two single-stranded 21mer RNA sequences: ‘21 R+’ (5'-AUGUGGAAAAUCUCUAGCAGU-3') and ‘21 R-’ (5'-CUGCUAGAGAUUUUCCACAU-3'). A short hairpin loop oligo (5’-GCTCTAGAGCATTATGTTCAGATAAGG-3’) was used to evaluate binding to small structured RNAs.(A) Representative membrane images of the bound and unbound signals. Filter binding experiments were conducted in experimental duplicates for each reaction. (B) Non-linear fits were used to generate binding curves for PepA and the three RNA substrates (‘21+’, “21- “, and ‘hairpin’). Error bars represent the variation between the experimental replicates. (C) Estimated KD values for each substrate.