Fig. 2

DMS analysis of MIT domain binding peptides and validation of predicted binding peptides from known interactor. (A, B) Heatmap representation of the PSSMs generated by the peptide-phage display-based DMS analysis of the two indicated peptides. (C) SPOT array results of selected peptides from known interactors tested for binding to the MIT domain. The highest scoring USP8 MIT binding peptides based on peptide array analysis of 24 predicted ligands are shown. The amino acid residues of the predicted motifs are indicated in bold. The DBP_{116 − 131} peptide was used a positive control, and the signal intensities were normalized to the highest intensity observed (that is, for DNAJB6) and indicated as percentage of max signal. The SPOT array and additional information can be found in Supplementary Fig. S4 and Supplementary Table S4. (D) SPOT array alanine scanning of DNAJB6_{118 − 130} peptide binding to the MIT domain. (E) SPOT array alanine scanning of CDH1_{863 − 877} peptide binding to the MIT domain. (D, E) Signal intensities were normalized to wild type (Wt) and displayed as average percent signal change