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An engineered cereblon optimized for high throughput screening and molecular glue discovery.

Cell chemical biology

Authors: Henry J Bailey, Jonathan Eisert, Rubina Kazi, Jan Gerhartz, Dominika Ewa Pieńkowska, Ina Dressel, Joshua Vollrath, Ivan Kondratov, Tetiana Matviyuk, Nataliya Tolmachova, Varun Jayeshkumar Shah, Giulio Giuliani, Thorsten Mosler, Thomas M Geiger, Ana M Esteves, Sandra P Santos, Raquel L Sousa, Tiago M Bandeiras, Eva-Maria Leibrock, Ulrike Bauer, Birgitta Leuthner, Julian D Langer, Ansgar A Wegener, Radosław P Nowak, Fiona J Sorrell, Ivan Dikic

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.

Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.

PMID: 39610248

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