Figure 6. Residue-specific mapping of RNC interactions.
(a) An overlay of 1H-15N correlation spectra (recorded at a 1H frequency of 950 MHz) of FLN5+31 RNC (black) and unfolded, isolated FLN5∆12 (red) highlighting resonances that are significantly broadened in the RNC. (b) Relative intensities of 1H-15N resonances of folded FLN5 (5 µM) and unfolded FLN5 Y719E (8 µM) in the presence of 1 molar equivalent of 70S ribosomes. (c) Relative intensities of FLN5+21, FLN5+31, FLN5+42, FLN5+67 and FLN5+110 RNCs as compared to a reference, made up of a composite consisting of FLN5∆12 and FLN5 Y719E, to monitor unfolded FLN5 (red), and FLN5-6∆18 to monitor unfolded FLN6 (green), and folded FLN5 from 1H-13C correlation spectra (cyan). A 5-point moving average is plotted as a guide; errors derived from spectral noise, n=1. The grey shaded area denotes occluded residues inaccessible to PEGylation.