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Purification of Ubiquitinated p53 Proteins from Mammalian Cells
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Purification of Ubiquitinated p53 Proteins from Mammalian Cells

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Targeted Degradation of 53BP1 Using Ubiquitin Variant Induced Proximity

Bayonle Aminu1, Julia Fux1, Evan Mallette1

  • 1Department of Molecular and Cellular Biology, College of Biological Science, University of Guelph, 50 Stone Rd E, Guelph, ON N1G 2W1, Canada.

Biomolecules
|April 23, 2022
PubMed

Insights

Researchers developed Ubiquitin Variant Induced Proximity (UbVIP) technology to target proteins for degradation using the ubiquitin-proteasome system (UPS). This novel approach successfully degraded the 53BP1 protein, offering potential for new therapeutics against diseases like cancer.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Proteomics

Background:

  • The ubiquitin-proteasome system (UPS) is crucial for protein degradation and targeted by therapeutics for diseases like cancer.
  • Limited utilization of ~600 human E3 ligases hinders therapeutic development, leaving many protein-protein interaction surfaces unexplored.

Purpose of the Study:

  • To develop a novel protein-based technology for targeted protein degradation using engineered E3 ligases.
  • To demonstrate the efficacy of Ubiquitin Variant Induced Proximity (UbVIP) in degrading specific target proteins.

Main Methods:

  • Structure-based protein engineering was used to create multi-domain fusion proteins (UbVIPs).
  • Synthetic Ubiquitin Variant (UbV) binders were generated to recruit E3 ligases.
  • A library of UbVIPs was designed to target the DNA damage response protein 53BP1.

Main Results:

  • Two UbVIPs, recruiting RFWD3 and NEDD4L E3 ligases, effectively induced proteasomal degradation of 53BP1 in human cell lines.
  • This demonstrated UbV technology's capability for targeted degradation of nuclear proteins.
  • The study provides a proof-of-concept for UbV technology in targeted protein degradation.

Conclusions:

  • UbV technology offers a versatile platform for developing protein-based targeted degradation therapeutics.
  • This approach can facilitate the identification and utilization of novel E3 ligases for therapeutic applications.
  • UbVIP technology shows promise for treating cancers and neurodegenerative disorders by enabling targeted protein destruction.
Keywords:
53BP1E3 ligaseNEDD4LPROTACRFWD3Ubiquitin Varianttargeted protein degradation

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