Integrating fragment-based screening with targeted protein degradation and genetic rescue to explore eIF4E function
Eukaryotic initiation factor 4E (eIF4E) serves as a regulatory hub for oncogene-driven protein synthesis and is considered a promising anticancer target. Here we screen a fragment library against eIF4E and identify a ligand-binding site with previously unknown function. Follow-up structure-based design yields a low nM tool compound (4, K<sub>d</sub> = 0.09 µM; LE 0.38), which disrupts the eIF4E:eIF4G interaction, inhibits translation in cell lysates, and demonstrates target engagement with eIF4E in intact cells (EC<sub>50</sub> = 2 µM). By coupling targeted protein degradation with genetic rescue using eIF4E mutants, we show that disruption of both the canonical eIF4G and non-canonical binding sites is likely required to drive a strong cellular effect. This work highlights the power of fragment-based drug discovery to identify pockets in difficult-to-drug proteins and how this approach can be combined with genetic characterization and degrader technology to probe protein function in complex biological systems.
Authors
- Marianna Martella (ORCID: https://orcid.org/0000-0003-2389-3975)
- Hugh Walton
- Andrew Woodhead (ORCID: https://orcid.org/0009-0005-1604-2904)
- M. Vinković
- Puja Pathuri
- C. Martinez-Fleites
- Sabrina D’Agostino (ORCID: https://orcid.org/0000-0002-4858-8873)
- Paul Andrew Clarke (ORCID: https://orcid.org/0000-0001-9342-1290)
- Steven D. Hiscock
- Marissa Powers (ORCID: https://orcid.org/0000-0003-0775-4908)
- Charlotte East
- Nick Palmer (ORCID: https://orcid.org/0009-0002-6095-7763)
- Elisabetta Chiarparin (ORCID: https://orcid.org/0000-0002-2998-1346)
- Kate Swabey
- Maria G. Carr
- Jeffrey D. St. Denis (ORCID: https://orcid.org/0000-0002-8653-6644)
- Paul N. Mortenson (ORCID: https://orcid.org/0000-0002-3583-3163)
- Christopher I. Milton (ORCID: https://orcid.org/0000-0002-1744-6071)
- George Ward (ORCID: https://orcid.org/0000-0003-3184-2211)
- Benjamin D. Cons (ORCID: https://orcid.org/0000-0001-9670-6670)
- Glyn Williams (ORCID: https://orcid.org/0000-0003-3199-4928)
- Caroline Richardson (ORCID: https://orcid.org/0000-0002-1305-4134)
- Susanne M Saalau (ORCID: https://orcid.org/0009-0008-3130-4521)
- Joseph E. Coyle
- Swee Y. Sharp
Institutions
- Institute of Cancer Research (GB)
- GW Pharmaceuticals (United Kingdom) (GB)
Publication Details
- Journal
- Nature Communications
- Published
- 2024-11-20
- DOI
- https://doi.org/10.1038/s41467-024-54356-1
- Citations
- 4
- Primary Topic
- Monoclonal and Polyclonal Antibodies Research
- Type
- article
- Field-Weighted Citation Impact
- 1.62
Funders
- Astex Pharmaceuticals