An engineered cereblon optimized for high-throughput screening and molecular glue discovery
The majority of clinical degraders utilize an immunomodulatory imide drug (IMiD)-based derivative that directs their target to the E3 ligase receptor cereblon (CRBN); however, identification of IMiD molecular glue substrates has remained underexplored. To tackle this, we design human CRBN constructs, which retain all features for ternary complex formation, while allowing generation of homogenous and cost-efficient expression in E. coli. Extensive profiling of the construct shows it to be the "best of both worlds" in terms of binding activity and ease of production. We next designed the "Enamine focused IMiD library" and demonstrated applicability of the construct to high-throughput screening, identifying binders with high potency, ligand efficiency, and specificity. Finally, we adapt our construct for proof of principle glue screening approaches enabling IMiD cellular interactome determination. Coupled with our IMiD binding landscape the methods described here should serve as valuable tools to assist discovery of next generation CRBN glues.
Authors
- Sandra P. Santos (ORCID: https://orcid.org/0000-0001-9153-5550)
- Thomas Geiger (ORCID: https://orcid.org/0000-0002-6278-3990)
- Ansgar Wegener (ORCID: https://orcid.org/0009-0005-5737-7460)
- Tetiana Matviyuk
- Nataliya A. Tolmachova
- F.J. Sorrell (ORCID: https://orcid.org/0000-0001-9211-5590)
- Joshua Vollrath (ORCID: https://orcid.org/0000-0002-6673-3050)
- Thorsten Mosler (ORCID: https://orcid.org/0000-0003-1800-2680)
- Julian D. Langer (ORCID: https://orcid.org/0000-0002-5190-577X)
- Eva-Maria Leibrock
- Rui Dinis Sousa (ORCID: https://orcid.org/0000-0001-7607-555X)
- Dominika Ewa Pieńkowska
- Ivan S. Kondratov (ORCID: https://orcid.org/0000-0003-0192-0725)
- Rubina Kazi
- Tiago M. Bandeiras (ORCID: https://orcid.org/0000-0001-6089-6089)
- Jonathan Eisert (ORCID: https://orcid.org/0009-0001-6850-5908)
- Birgitta Leuthner (ORCID: https://orcid.org/0009-0000-5476-7854)
- Radosław P. Nowak (ORCID: https://orcid.org/0000-0002-0605-0071)
- Ivan Đikić (ORCID: https://orcid.org/0000-0001-8156-9511)
- Giulio Giuliani (ORCID: https://orcid.org/0000-0002-4648-0076)
- Ulrike Bauer (ORCID: https://orcid.org/0000-0002-4701-793X)
- Jan Gerhartz (ORCID: https://orcid.org/0009-0002-9457-4656)
- Ana M. Esteves
- H. Bailey (ORCID: https://orcid.org/0000-0002-9040-0395)
- Varun Jayeshkumar Shah
- Ina Dressel
Institutions
- Goethe University Frankfurt (DE)
- University of Bonn (DE)
- Merck KGaA, Darmstadt (Germany) (DE)
- Frankfurt Institute for Advanced Studies (DE)
- Institute of Bioorganic Chemistry and Petrochemistry V.P. Kukhar (UA)
- Max Planck Institute of Biophysics (DE)
- Max Planck Institute for Brain Research (DE)
- Instituto de Biologia Experimental e Tecnológica (PT)
- Enamine (Ukraine) (UA)
Publication Details
- Journal
- Cell chemical biology
- Published
- 2024-11-27
- DOI
- https://doi.org/10.1016/j.chembiol.2024.11.002
- Citations
- 19
- Primary Topic
- Protein Degradation and Inhibitors
- Type
- article
- Field-Weighted Citation Impact
- 4.44
Funders
- Deutsche Forschungsgemeinschaft
- Bundesministerium für Bildung und Forschung