2. Academic Validation
  3. Design of a Cereblon construct for crystallographic and biophysical studies of protein degraders

Design of a Cereblon construct for crystallographic and biophysical studies of protein degraders

  • Nat Commun. 2024 Oct 15;15(1):8885. doi: 10.1038/s41467-024-52871-9.
Alena Kroupova 1,  Valentina A Spiteri 1,  Zoe J Rutter 1,  Hirotake Furihata 1,  Darren Darren 1 2,  Sarath Ramachandran 1 3,  Sohini Chakraborti 1,  Kevin Haubrich 1,  Julie Pethe 1 4,  Denzel Gonzales 1 5,  Andre J Wijaya 1 6,  Maria Rodriguez-Rios 1,  Manon Sturbaut 1,  Dylan M Lynch 1,  William Farnaby 1,  Mark A Nakasone 1,  David Zollman 7,  Alessio Ciulli 8
Affiliations

Affiliations

  • 1 Centre for Targeted Protein Degradation, School of Life Sciences, University of Dundee, Dundee, DD1 5JJ, UK.
  • 2 Cancer Science Institute Singapore, National University of Singapore, Singapore, 117599, Singapore.
  • 3 Biocon BMS R&D Center, Bommasandra Industrial Area, Bommasandra, Karnataka, 560099, India.
  • 4 National Heart and Lung Institute, Imperial College London, London, SW3 6LY, UK.
  • 5 Institute of Cell Biology, University of Edinburgh, Edinburgh, EH9 3BF, UK.
  • 6 PT Kalbe Farma, Jl. Let. Jend Suprapto Kav 4, Kalbe Farma, Jakarta, 10510, Indonesia.
  • 7 Centre for Targeted Protein Degradation, School of Life Sciences, University of Dundee, Dundee, DD1 5JJ, UK. d.l.zollman@dundee.ac.uk.
  • 8 Centre for Targeted Protein Degradation, School of Life Sciences, University of Dundee, Dundee, DD1 5JJ, UK. a.ciulli@dundee.ac.uk.
PMID: 39406745 DOI: 10.1038/s41467-024-52871-9
Abstract

The ubiquitin E3 ligase Cereblon (CRBN) is the target of therapeutic drugs thalidomide and lenalidomide and is recruited by most targeted protein degraders (PROTACs and Molecular Glues) in clinical development. Biophysical and structural investigation of CRBN has been limited by current constructs that either require co-expression with the adaptor DDB1 or inadequately represent full-length protein, with high-resolution structures of degrader ternary complexes remaining rare. We present the design of CRBNmidi, a construct that readily expresses from E. coli with high yields as soluble, stable protein without DDB1. We benchmark CRBNmidi for wild-type functionality through a suite of biophysical techniques and solve high-resolution co-crystal structures of its binary and ternary complexes with degraders. We qualify CRBNmidi as an enabling tool to accelerate structure-based discovery of the next generation of CRBN based therapeutics.

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