E3-Specific Degrader Discovery by Dynamic Tracing of Substrate Receptor Abundance

Targeted protein degradation (TPD) is a new pharmacology based on small-molecule degraders that induce proximity between a protein of interest (POI) and an E3 ubiquitin ligase. Of the approximately 600 E3s encoded in the human genome, only around 2% can be co-opted with degraders. This underrepresentation is caused by a paucity of discovery approaches to identify degraders for defined E3s. This hampers a rational expansion of the druggable proteome and stymies critical advancements in the field, such as tissue-and cell-specific degradation. Here, we focus on dynamic NEDD8 conjugation, a post-translational, regulatory circuit that controls the activity of 250 cullin RING E3 ligases (CRLs). Leveraging this regulatory layer enabled us to develop a scalable assay to identify compounds that alter the interactome of an E3 of interest by tracing their abundance after pharmacologically induced auto-degradation. Initial validation studies are performed for CRBN and VHL, but proteomics studies indicate broad applicability for many CRLs. Among amenable ligases, we select CRLDCAF15 for a proof-of-concept screen, leading to the identification of a novel DCAF15-dependent molecular glue degrader inducing the degradation of RBM23 and RBM39. Together, this strategy empowers the scalable identification of degraders specific to a ligase of interest.

Automated summary

Targeted protein degradation (TPD) is a new pharmacology that uses small-molecule degraders to induce proximity between a protein of interest (POI) and an E3 ubiquitin ligase. Only a small percentage of E3s can be co-opted with degraders due to a lack of discovery approaches. In this study, the focus is on dynamic NEDD8 conjugation, which enables the identification of compounds that alter the interactome of a specific E3 ligase. This strategy allows for scalable identification of degraders specific to a ligase of interest.