2. Academic Validation
  3. A chemical probe to modulate human GID4 Pro/N-degron interactions

A chemical probe to modulate human GID4 Pro/N-degron interactions

  • Nat Chem Biol. 2024 May 21. doi: 10.1038/s41589-024-01618-0.
Dominic D G Owens # 1,  Matthew E R Maitland # 1 2 3 4,  Aliakbar Khalili Yazdi 1,  Xiaosheng Song 1,  Viviane Reber 5,  Martin P Schwalm 6 7,  Raquel A C Machado 1,  Nicolas Bauer 6 7,  Xu Wang 2,  Magdalena M Szewczyk 1,  Cheng Dong 1,  Aiping Dong 1,  Peter Loppnau 1,  Matthew F Calabrese 8,  Matthew S Dowling 8,  Jisun Lee 8,  Justin I Montgomery 8,  Thomas N O'Connell 8,  Chakrapani Subramanyam 8,  Feng Wang 8,  Ella C Adamson 1,  Matthieu Schapira 1 9,  Matthias Gstaiger 5,  Stefan Knapp 6 7,  Masoud Vedadi 1 9,  Jinrong Min 1 10 11,  Gilles A Lajoie 3 4,  Dalia Barsyte-Lovejoy 1 9,  Dafydd R Owen 8,  Caroline Schild-Poulter 2 3,  Cheryl H Arrowsmith 12 13 14
Affiliations

Affiliations

  • 1 Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada.
  • 2 Robarts Research Institute, Schulich School of Medicine and Dentistry, University of Western Ontario, London, Ontario, Canada.
  • 3 Department of Biochemistry, Schulich School of Medicine and Dentistry, University of Western Ontario, London, Ontario, Canada.
  • 4 Don Rix Protein Identification Facility, Schulich School of Medicine and Dentistry, University of Western Ontario, London, Ontario, Canada.
  • 5 Institute of Molecular Systems Biology at ETH Zurich, Zurich, Switzerland.
  • 6 Institut für Pharmazeutische Chemie, Goethe-University Frankfurt, Biozentrum, Frankfurt am Main, Germany.
  • 7 Structural Genomics Consortium, Goethe-University Frankfurt, Buchmann Institute for Life Sciences, Frankfurt am Main, Germany.
  • 8 Development and Medical, Pfizer Worldwide Research, Groton, CT, USA.
  • 9 Department of Pharmacology and Toxicology, University of Toronto, Toronto, Ontario, Canada.
  • 10 Department of Physiology, University of Toronto, Toronto, Ontario, Canada.
  • 11 Hubei Key Laboratory of Genetic Regulation and Integrative Biology, School of Life Sciences, Central China Normal University, Wuhan, China.
  • 12 Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada. Cheryl.Arrowsmith@uhn.ca.
  • 13 Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada. Cheryl.Arrowsmith@uhn.ca.
  • 14 Princess Margaret Cancer Centre, University Health Network, Toronto, Ontario, Canada. Cheryl.Arrowsmith@uhn.ca.
  • # Contributed equally.
PMID: 38773330 DOI: 10.1038/s41589-024-01618-0
Abstract

The C-terminal to LisH (CTLH) complex is a ubiquitin Ligase complex that recognizes substrates with Pro/N-degrons via its substrate receptor Glucose-Induced Degradation 4 (GID4), but its function and substrates in humans remain unclear. Here, we report PFI-7, a potent, selective and cell-active chemical probe that antagonizes Pro/N-degron binding to human GID4. Use of PFI-7 in proximity-dependent biotinylation and quantitative proteomics enabled the identification of GID4 interactors and GID4-regulated proteins. GID4 interactors are enriched for nucleolar proteins, including the Pro/N-degron-containing RNA helicases DDX21 and DDX50. We also identified a distinct subset of proteins whose cellular levels are regulated by GID4 including HMGCS1, a Pro/N-degron-containing metabolic Enzyme. These data reveal human GID4 Pro/N-degron targets regulated through a combination of degradative and nondegradative functions. Going forward, PFI-7 will be a valuable research tool for investigating CTLH complex biology and facilitating development of targeted protein degradation strategies that highjack CTLH E3 Ligase activity.

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