2. Academic Validation
  3. Continuous evolution of compact protein degradation tags regulated by selective molecular glues

Continuous evolution of compact protein degradation tags regulated by selective molecular glues

  • Science. 2024 Mar 15;383(6688):eadk4422. doi: 10.1126/science.adk4422.
Jaron A M Mercer # 1 2 3,  Stephan J DeCarlo # 1 2 3,  Shourya S Roy Burman # 4 5,  Vedagopuram Sreekanth 6 7 8,  Andrew T Nelson 1 2 3,  Moritz Hunkeler 4 5,  Peter J Chen 1 2 3,  Katherine A Donovan 4 5,  Praveen Kokkonda 6 7,  Praveen K Tiwari 6 7 8,  Veronika M Shoba 6 7,  Arghya Deb 6 7,  Amit Choudhary 6 7 8,  Eric S Fischer 4 5,  David R Liu 1 2 3
Affiliations

Affiliations

  • 1 Merkin Institute of Transformative Technologies in Healthcare, Broad Institute of Harvard and MIT, Cambridge, MA 02142, USA.
  • 2 Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA 02138, USA.
  • 3 Howard Hughes Medical Institute, Harvard University, Cambridge, MA 02138, USA.
  • 4 Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, MA 02215, USA.
  • 5 Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA.
  • 6 Chemical Biology and Therapeutics Science, Broad Institute of Harvard and MIT, Cambridge, MA 02142, USA.
  • 7 Department of Medicine, Harvard Medical School, Boston, MA 02115, USA.
  • 8 Divisions of Renal Medicine and Engineering, Brigham and Women's Hospital, Boston, MA 02115, USA.
  • # Contributed equally.
PMID: 38484051 DOI: 10.1126/science.adk4422
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.

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