From protein-protein interaction to E2 inhibitors leads: identification of peptide binding of UbcH10

Ubiquitin Conjugating Enzyme UbcH10, is a member of Ubiquitin Conjugation Enzymes (Ubc) family, involved in the regulation of cell cycle, and, when functionally altered, in cancer pathologies.[1,2] Here we present the de-novo design, the affinity assays and molecular dynamics studies of the binding mode of two peptide as candidate inhibitors of UbcH10-partner interaction. On the basis of the computational model of the quaternary complex between hUbA1, two ubiquitin molecules and the E2 enzyme, UbcH10, we have designed 3 families of 13-mer inhibitory peptides potentially able to impair UbcH10-partner interaction and UbcH10 activity. These peptides were synthetised biotinylated and subjected to affinity assays (ELISA) which allowed us to identify two potential leads, bearing to two different families, showing a sequence-dependent binding with micromolar affinity toward UbcH10. The binding mode of candidate leads was analysed by a computational study including peptide-protein docking and extensive molecular dynamics calculations.