Work

Site-resolved assessment of targeted protein degradation

Year: 2025

Type: article

Abstract: Induced proximity using small molecules, exemplified by targeted protein degradation (TPD), represents a highly promising therapeutic strategy with significant untapped potential. However, evaluating an induced proximity event that accurately reflects drug binding typically requires the challenging and costly development of specific ligands, which limits the advancement of medicines based on this modality. To overcome this bottleneck, we combine genetic code expansion with ultra-fast bioorthogonal chemistry to sensitize specific protein sites at single-residue resolution to a generic bioorthogonal proximity inducer (BPI) molecule. Mammalian cells expressing sensitized mutants of the ubiquitin E3 ligases VHL and CRBN exhibit neosubstrate degradation in the presence of a BPI equipped with a ligand targeting bromodomain and extraterminal (BET) proteins. Furthermore, we demonstrate E3-independent degradation through recruitment of an upstream E2 conjugating enzyme. We anticipate that this approach will have broad applicability, enabling comprehensive assessment of the scope of induced proximity.

Source: Cell chemical biology

DOI: 10.1016/j.chembiol.2025.06.002

Authors: Ricardo Moreno-Ballesteros, Thomas Pembridge, Gaurav Beniwal, Satpal Virdee

Institutions: University of Dundee, MRC Protein Phosphorylation and Ubiquitylation Unit

Language: English

Topic: Protein Degradation and Inhibitors

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: hybrid

Estimated APC paid: $9,350

Funders: Scottish Enterprise, Vienna Science and Technology Fund, Medical Research Council