Fig. 3
Overview of USP8 Rhod domain binding to ligands identified through ProP-PD selections. (A) Peptides of high/medium confidence binding to USP8 Rhodanese domain identified in ProP-PD. (B) Fluorescence polarization-monitored experiments of TET31705 − 1720 bound to the USP8 Rhod domain, displaced by the peptide ligands KIAA1614568 − 583 and TET31705 − 1720. The data are presented as mean ± SD (N = 3 technical triplicates) and the KD values are indicated. (C) Peptide SPOT array alanine scanning of KIAA1614568 − 583 peptide binding to the Rhod domain. (D) Peptide SPOT array alanine scanning of TET31705 − 1720 peptide binding to the Rhod domain. (C, D) Amino acid residues disrupting binding when mutated to alanine are shown in bold. Signal intensities were normalized to wild type (Wt) and displayed as average percent signal change. (E) AF3 model of TET31705 − 1720 peptide (teal) binding to the Rhod domain. Amino acids of the USP8 Rhod domain that interact with the peptide based on the model are shown in yellow