USP7-CTD interacts with GMPS and UHRF1 peptides.

(A) Alignment and dissociation constants of ICP0, GMPS and UHRF1 peptides. (B) Location of the KxxxK motif (³¹⁶DRTPRKRISKTLN³²⁸) within a disordered loop on the GMPS crystal structure (PDB ID 2VXO). (C) Coomassie stained gel of GST pull-down assays with WT or mutant USP7-CTD (D762A, D764A and D762A/D764A) and GST-GMPS peptide. Lane 1 USP7-CTD and GST-GMPS peptide load, lanes 2–5 are the eluted fractions with WT and mutant USP7-CTD. (D) Coomassie stained gel of GST pull-down assays with WT USP7-CTD and mutant GST-GMPS peptides. Lanes 1 (load) and 2 (elute) USP7-CTD and WT GST-GMPS, lanes 3–5 are the eluted fractions with GST-GMPS mutants (K321A, K325A and K321A/K325A). (E) Coomassie stained gel of GST pull-down assays with WT or mutant USP7-CTD (D762A, D762A/D764A and D762R/D764R) and GST-UHRF1 peptide. Lanes 1–4 are the loaded fractions with WT and mutant USP7-CTD. Lanes 5–8 are the eluted fractions with WT and mutant USP7-CTD. (F) Coomassie stained gel of GST pull-down assays with WT USP7-CTD and mutant GST-UHRF1 peptides. Lanes 1–4 are the loaded fractions with WT and mutant (K644A, K648A and K644A/K648A) GST-UHRF1. Lanes 5–8 are the eluted fractions with WT and mutant (K644A, K648A and K644A/K648A) GST-UHRF1. In all instances, approximately 1–2% of the input and 2.5% of the eluate is loaded on the gels.