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Regulated Protein Degradation

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It is vital to regulate the activity of enzymatic as well as non-enzymatic proteins inside the cell. This can be achieved either through creating a balance between their rate of synthesis and degradation or regulating the intrinsic activity of the protein. Both these regulation mechanisms play an essential role in the normal functioning of cells.
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Updated: Dec 10, 2025

Using Phage Display to Develop Ubiquitin Variant Modulators for E3 Ligases
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Using Phage Display to Develop Ubiquitin Variant Modulators for E3 Ligases

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Targeted modulation of E3 ligases using engineered ubiquitin variants.

Nicole LeBlanc1, Evan Mallette1, Wei Zhang1,2

  • 1Department of Molecular and Cellular Biology, College of Biological Science, University of Guelph, Guelph, ON, Canada.

The FEBS Journal
|September 1, 2020
PubMed
Summary

Researchers engineered ubiquitin variants (UbVs) to create novel inhibitors and activators for E3 ubiquitin ligases, crucial regulators of cellular processes and disease. This protein engineering approach offers a powerful toolkit for modulating E3 ligase activity in drug discovery and research.

Keywords:
E3 ligasesactivatorcancerinhibitorphage displayprotein engineeringsignal transductionubiquitin variantubiquitination

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In Vitro Analysis of E3 Ubiquitin Ligase Function
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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cellular Biology

Background:

  • Ubiquitination is a key post-translational modification regulating cellular processes.
  • E3 ubiquitin ligases are critical determinants of ubiquitination specificity and are implicated in human diseases like cancer.
  • Targeting E3 ligase activity is essential for disease research and drug discovery.

Purpose of the Study:

  • To summarize advancements in using protein engineering to develop ubiquitin variant (UbV) based modulators for E3 ligases.
  • To present UbV inhibitors for all major E3 ligase families.
  • To introduce UbV activators for specific E3 ligase types.

Main Methods:

  • Protein engineering of ubiquitin variants (UbVs).
  • Development of UbV-based inhibitors targeting diverse E3 ligase families.
  • Design of UbV-based activators for homologous with E6-associated protein C terminus (HECT) E3s and homodimeric RING E3s.

Main Results:

  • Successful generation of UbV inhibitors applicable to a broad range of E3 ligases.
  • Development of UbV activators for HECT and homodimeric RING E3s.
  • Demonstration of a general strategy for modulating E3 ligase activity.

Conclusions:

  • UbV-based molecules provide a versatile toolkit for inhibiting or activating E3 ligases.
  • These synthetic molecules hold significant potential for exploring protein degradation pathways in drug discovery.
  • The protein engineering approach offers a promising avenue for therapeutic development targeting E3 ligase dysfunction.