Fig. 3

In vitro acetylation competition assay. a Schematic representation of the in vitro competition acetylation assay. b Heat map displaying in vitro acylation specificities of HATs during acyl-CoA competition assays. Different HATs were assayed against histones H3 or H4 as specified in the table headers in the presence of equimolar concentrations of acetyl-CoA and a competing acyl-CoA donor. Negative controls with no enzyme are also shown. c Correlation between the molecular weight and the acylation preference for different acyl donors displayed for the HATs GCN5 on histone H3, d p300 on histone H3 and e MOF on histone H4. For each modification, molecular weight is indicated. Results show that the preference for acetyl-CoA over the other acyl donor tightly correlates with the molecular weight of the acyl donor. Crotonylation was not included in the correlations, as its molecular weight did not correlate well with the preference of the enzymes over acetyl-CoA. All graphs are shown as the average of log2 ratios between the relative abundances of all acetylated peptides and the relative abundances of the corresponding competing acylated peptide