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

Autor: Owens DDG; Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada., Maitland MER; Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada.; Robarts Research Institute, Schulich School of Medicine and Dentistry, University of Western Ontario, London, Ontario, Canada.; Department of Biochemistry, Schulich School of Medicine and Dentistry, University of Western Ontario, London, Ontario, Canada.; Don Rix Protein Identification Facility, Schulich School of Medicine and Dentistry, University of Western Ontario, London, Ontario, Canada., Khalili Yazdi A; Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada., Song X; Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada., Reber V; Institute of Molecular Systems Biology at ETH Zurich, Zurich, Switzerland., Schwalm MP; Institut für Pharmazeutische Chemie, Goethe-University Frankfurt, Biozentrum, Frankfurt am Main, Germany.; Structural Genomics Consortium, Goethe-University Frankfurt, Buchmann Institute for Life Sciences, Frankfurt am Main, Germany., Machado RAC; Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada., Bauer N; Institut für Pharmazeutische Chemie, Goethe-University Frankfurt, Biozentrum, Frankfurt am Main, Germany.; Structural Genomics Consortium, Goethe-University Frankfurt, Buchmann Institute for Life Sciences, Frankfurt am Main, Germany., Wang X; Robarts Research Institute, Schulich School of Medicine and Dentistry, University of Western Ontario, London, Ontario, Canada., Szewczyk MM; Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada., Dong C; Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada., Dong A; Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada., Loppnau P; Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada., Calabrese MF; Development and Medical, Pfizer Worldwide Research, Groton, CT, USA., Dowling MS; Development and Medical, Pfizer Worldwide Research, Groton, CT, USA., Lee J; Development and Medical, Pfizer Worldwide Research, Groton, CT, USA., Montgomery JI; Development and Medical, Pfizer Worldwide Research, Groton, CT, USA., O'Connell TN; Development and Medical, Pfizer Worldwide Research, Groton, CT, USA., Subramanyam C; Development and Medical, Pfizer Worldwide Research, Groton, CT, USA., Wang F; Development and Medical, Pfizer Worldwide Research, Groton, CT, USA., Adamson EC; Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada., Schapira M; Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada.; Department of Pharmacology and Toxicology, University of Toronto, Toronto, Ontario, Canada., Gstaiger M; Institute of Molecular Systems Biology at ETH Zurich, Zurich, Switzerland., Knapp S; Institut für Pharmazeutische Chemie, Goethe-University Frankfurt, Biozentrum, Frankfurt am Main, Germany.; Structural Genomics Consortium, Goethe-University Frankfurt, Buchmann Institute for Life Sciences, Frankfurt am Main, Germany., Vedadi M; Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada.; Department of Pharmacology and Toxicology, University of Toronto, Toronto, Ontario, Canada., Min J; Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada.; Department of Physiology, University of Toronto, Toronto, Ontario, Canada.; Hubei Key Laboratory of Genetic Regulation and Integrative Biology, School of Life Sciences, Central China Normal University, Wuhan, China., Lajoie GA; Department of Biochemistry, Schulich School of Medicine and Dentistry, University of Western Ontario, London, Ontario, Canada.; Don Rix Protein Identification Facility, Schulich School of Medicine and Dentistry, University of Western Ontario, London, Ontario, Canada., Barsyte-Lovejoy D; Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada.; Department of Pharmacology and Toxicology, University of Toronto, Toronto, Ontario, Canada., Owen DR; Development and Medical, Pfizer Worldwide Research, Groton, CT, USA., Schild-Poulter C; Robarts Research Institute, Schulich School of Medicine and Dentistry, University of Western Ontario, London, Ontario, Canada.; Department of Biochemistry, Schulich School of Medicine and Dentistry, University of Western Ontario, London, Ontario, Canada., Arrowsmith CH; Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada. Cheryl.Arrowsmith@uhn.ca.; Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada. Cheryl.Arrowsmith@uhn.ca.; Princess Margaret Cancer Centre, University Health Network, Toronto, Ontario, Canada. Cheryl.Arrowsmith@uhn.ca.
Jazyk: angličtina
Zdroj: Nature chemical biology [Nat Chem Biol] 2024 Sep; Vol. 20 (9), pp. 1164-1175. Date of Electronic Publication: 2024 May 21.
DOI: 10.1038/s41589-024-01618-0
Abstrakt: 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.
(© 2024. The Author(s), under exclusive licence to Springer Nature America, Inc.)
Databáze: MEDLINE
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