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Title: Inhibition of SCF ubiquitin ligases by engineered ubiquitin variants that target the Cul1 binding site on the Skp1–F-box interface

Journal Article · · Proceedings of the National Academy of Sciences of the United States of America
 [1];  [2];  [1];  [2];  [1];  [3];  [1];  [4];  [1];  [5];  [1]
  1. Univ. of Toronto, ON (Canada). Terrence Donnelly Center for Cellular and Biomolecular Research. Banting and Best Dept. of Medical Research. Dept. of Molecular Genetics
  2. Mount Sinai Hospital, Toronto, ON (Canada). Lunenfeld-Tanenbaum Research Inst.
  3. Cornell Univ., Argonne, IL (United States). Dept. of Chemistry and Chemical Biology
  4. Mount Sinai Hospital, Toronto, ON (Canada). Lunenfeld-Tanenbaum Research Inst.; Univ. of Montreal, QC (Canada). Inst. of Immunology and Cancer Research
  5. Mount Sinai Hospital, Toronto, ON (Canada). Lunenfeld-Tanenbaum Research Inst.; Univ. of Toronto, ON (Canada). Dept. of Molecular Genetics
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Skp1–Cul1–F-box (SCF) E3 ligases play key roles in multiple cellular processes through ubiquitination and subsequent degradation of substrate proteins. Although Skp1 and Cul1 are invariant components of all SCF complexes, the 69 different human F-box proteins are variable substrate binding modules that determine specificity. SCF E3 ligases are activated in many cancers and inhibitors could have therapeutic potential. Here, we used phage display to develop specific ubiquitin-based inhibitors against two F-box proteins, Fbw7 and Fbw11. Unexpectedly, the ubiquitin variants bind at the interface of Skp1 and F-box proteins and inhibit ligase activity by preventing Cul1 binding to the same surface. Using structure-based design and phage display, we modified the initial inhibitors to generate broad-spectrum inhibitors that targeted many SCF ligases, or conversely, a highly specific inhibitor that discriminated between even the close homologs Fbw11 and Fbw1. We propose that most F-box proteins can be targeted by this approach for basic research and for potential cancer therapies.

Research Organization:
Univ. of Toronto, ON (Canada)
Sponsoring Organization:
USDOE Office of Science (SC), Basic Energy Sciences (BES); National Inst. of Health (NIH) (United States); Canadian Inst. of Health Research (CIHR) (Canada); Canadian Cancer Society Research Inst. (Canada)
Contributing Organization:
Mount Sinai Hospital, Toronto, ON (Canada); Cornell Univ., Argonne, IL (United States); Univ. of Montreal, QC (Canada)
Grant/Contract Number:
AC02-06CH11357; GM103403; MOP-136956; MOP-126129
OSTI ID:
1247362
Journal Information:
Proceedings of the National Academy of Sciences of the United States of America, Vol. 113, Issue 13; ISSN 0027-8424
Publisher:
National Academy of Sciences, Washington, DC (United States)Copyright Statement
Country of Publication:
United States
Language:
ENGLISH
Citation Metrics:
Cited by: 48 works
Citation information provided by
Web of Science

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Cited By (13)

Humanization and directed evolution of the selenium-containing scFv phage abzyme journal January 2018
Identification and Characterization of Mutations in Ubiquitin Required for Non-covalent Dimer Formation journal September 2019
UBB pseudogene 4 encodes functional ubiquitin variants journal March 2020
Proteomic identification of the oncoprotein STAT3 as a target of a novel Skp1 inhibitor journal November 2016
Specific targeting of the deubiquitinase and E3 ligase families with engineered ubiquitin variants: Gorelik and Sidhu journal November 2016
A General Strategy for Discovery of Inhibitors and Activators of RING and U-box E3 Ligases with Ubiquitin Variants journal October 2017
Saturation scanning of ubiquitin variants reveals a common hot spot for binding to USP2 and USP21 journal July 2016
Protein Engineering in the Ubiquitin System: Tools for Discovery and Beyond journal February 2020
Structure and Function of Viral Deubiquitinating Enzymes journal November 2017
Site-specific inhibition of the small ubiquitin-like modifier (SUMO)-conjugating enzyme Ubc9 selectively impairs SUMO chain formation journal August 2017
Peptides meet ubiquitin: Simple interactions regulating complex cell signaling journal September 2018
Emerging drug development technologies targeting ubiquitination for cancer therapeutics journal July 2019
Peptide design by optimization on a data-parameterized protein interaction landscape journal October 2018

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Related Subjects

60 APPLIED LIFE SCIENCES
59 BASIC BIOLOGICAL SCIENCES
Cul1 affinity
SCF inhibitors
Fbxw7
Fbxw11
β-Trcp