Establishment of a method to measure the interaction between nedd4 and UbCH5c for drug screening

Abstract

Protein ubiquitination is closely associated with tumorigenesis. E3 ubiquitin ligase Nedd4 is overexpressed in prostate, bladder, and colorectal cancer. Nedd4 interacts with UbCH5c ubiquitin transferase through its HECT domain. Two GST-tagged Nedd4 proteins and His-HA-UbCH5c protein are expressed and purified. Using these recombinant proteins, a method is established to measure the interaction between Nedd4 and UbCH5c. This established method can be used to screen the small molecule inhibitors to block the interaction between Nedd4 and UbCH5c, which inhibit the activity of E3 ubiquitin ligase Nedd4.