Effects of individual pseudouridine replacements on chemical shifts in the WT NSR bound to ribostamycin. Comparison of the imino proton 1H (A) and 31P (B) 1D NMR-spectra of the WT-NSR (top), the U14ψ variant (second row from top), the U13ψ variant (middle), the U18ψ variant (second row from bottom) and the U13ψ/U18ψ double replacement variant (bottom). The relevant signal assignments are given in the WT spectra (top). Assignments for pseudouridine H1 imino protons are given in blue. Relevant chemical shift differences between the WT and the pseudouridine containing variants are highlighted by red arrows. (C) Loop sequence of the WT NSR. Nucleotides that were replaced with pseudouridine are highlighted in bold italic. Nucleotides with relevant chemical shift differences in A and B are colored red.
