Fig. 4.
Structural insight into the selectivity of AF9-YEATS. a Alignment of the YEATS domain sequences: absolutely, moderately, and weakly conserved residues are colored green, orange, and pink, respectively. b Binding affinities of AF9-YEATS to indicated histone peptides. Values represent the average of three separate experiments with error calculated as the SD between the runs. c Structural overlay of the acyllysine-binding sites in the AF9-YEATS:H3K9bu (pink) and AF9-YEATS:H3K9cr17 (light gray) complexes. Yellow dashed lines represent short distances indicative of the aromatic-amide-aromatic π stacking interaction in AF9-YEATS:H3K9cr17. Red dashed lines represent short, <4 Å, distances between the crotonyl alkene group and the aromatic residues. d Superimposed 1H,15N HSQC spectra of AF9-YEATS, wt and Y78W mutant, recorded in the presence of increasing concentration of H3K9cr peptide. Spectra are color coded according to the protein:peptide molar ratio. e Representative binding curves used to determine Kd of AF9-YEATS Y78W by tryptophan fluorescence. f The ribbon diagram of the AF9-YEATS Y78W:H3K9cr complex. Red dashed lines represent short, <4 Å, distances between the crotonyl alkene group of H3K9cr and the aromatic residues of the protein