Fig. 6: Solid-state NMR reveals change in conformational dynamics of the histone H4 tail upon lysine 20 mono-methylation.
a Schematic representation of the AA sequence of human histone H4. The green flag highlights the position of AA K20 that is subject to methylation. Overlaid 2D 1H-13C refocused J-based INEPT spectra of 15-mer nucleosome arrays containing b H4K20me0 (black) and H4K20me1 (blue) and c H4K20me0 (black) and H4K20me3 (red) with uniformly 13C/15N labeled H4. The absence of K20 1Hα−13Cα correlations in the spectra (b, c) confirms that the K20me1 and K20me3 modifications of H4 were successful. d Zoomed-in plots of AA V21 showing the 1Hα−13Cα peaks of the three arrays. e AA site-specific peak intensities obtained from the two-dimensional (2D) NCA spectra (Supplementary Fig. 14). Peak intensities were normalized to the highest peak intensity of the NCA spectrum, error bars were derived from the RMSD value of the noises in the spectra. The NCA (correlation achieved by magnetization transfer from the N to alpha Cα) and the NCO (correlation achieved by magnetization transfer from the N to carbonyl CO) peak intensities report on the relative AA site-specific dynamic behavior. Here, only H4K20me1 (blue) or H4K20me3 (red) residues having considerable peak intensity differences compared with H4K20me0 (black) nucleosome arrays are displayed. The blue and red boxes highlight the residues exhibiting differences for the H4K20me1 and H4K20me3, respectively, in comparison with the H4K20me0 arrays. The complete peak intensity profiles are shown in Supplementary Fig. 15; the asterisk indicates that the peak intensity is not extracted because of overlapping peaks.
