Continuous evolution of compact protein degradation tags regulated by selective molecular glues.
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Abstract | Conditional protein degradation tags (degrons) are usually >100 amino acids long or are triggered by small molecules with substantial off-target effects, thwarting their use as specific modulators of endogenous protein levels. We developed a phage-assisted continuous evolution platform for molecular glue complexes (MG-PACE) and evolved a 36-amino acid zinc finger (ZF) degron (SD40) that binds the ubiquitin ligase substrate receptor cereblon in complex with PT-179, an orthogonal thalidomide derivative. Endogenous proteins tagged in-frame with SD40 using prime editing are degraded by otherwise inert PT-179. Cryo-electron microscopy structures of SD40 in complex with ligand-bound cereblon revealed mechanistic insights into the molecular basis of SD40's activity and specificity. Our efforts establish a system for continuous evolution of molecular glue complexes and provide ZF tags that overcome shortcomings associated with existing degrons. |
Year of Publication | 2024
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Journal | Science (New York, N.Y.)
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Volume | 383
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Issue | 6688
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Pages | eadk4422
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Date Published | 03/2024
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ISSN | 1095-9203
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DOI | 10.1126/science.adk4422
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PubMed ID | 38484051
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