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Updated: Oct 22, 2026

High-Throughput Cellular Profiling of Targeted Protein Degradation Compounds Using HiBiT CRISPR Cell Lines
Published on: November 9, 2020
Proteome-wide C-degron activity profiling connects conditional regulation of the CTLH E3 ligase complex to ribosome
Drew W Grant1, Shengjiang Tan2, Dylan E Ramage1
1Cambridge Institute for Therapeutic Immunology and Infectious Disease, Department of Medicine, University of Cambridge, Puddicombe Way, Cambridge CB2 0AW, UK.
Abstract:
Many E3 ubiquitin ligases recognize degrons located at protein termini, but the paucity of bona fide substrates of N-degron and C-degron pathways hampers our understanding of their physiological significance. Devising an expression screening strategy to assess the effect of C-terminal "capping" on the stability of thousands of human proteins, we systematically identify substrates harboring C-degrons. Interrogating one leading candidate, ZMYND19, we characterize a C-degron pathway governed via the C-terminal to LisH (CTLH) E3 ligase complex subunit Muskelin. Cell-to-cell variability in ZMYND19 stability uncovered conditional regulation, with CTLH-mediated degradation impaired by TNF-α stimulation but enhanced by mTOR inhibition. Parallel genetic and proteomic screens identified angio-associated migratory protein (AAMP) and apoptosis-enhancing nuclease (AEN) as additional substrates of the CTLHMuskelin C-degron pathway, leading us to define an essential role for AAMP in ribosome maturation through chaperone activity toward uL16. Altogether, these data define a C-degron pathway connecting conditional regulation of the CTLH E3 ligase complex to ribosome biogenesis.
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