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Updated: Jun 12, 2025
06:06
In Vitro Analysis of E3 Ubiquitin Ligase Function
Published on: May 14, 2021
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Targeted protein degradation using chimeric human E2 ubiquitin-conjugating enzymes.
Jonathan D Taylor1, Nathalie Barrett2, Sergio Martinez Cuesta3
1Biologics Engineering, R&D Oncology, AstraZeneca, Cambridge, CB2 0AA, UK. jonathan.taylor@astrazeneca.com.
Communications Biology
|September 19, 2024
Summary
Researchers developed novel E2 bioPROTACs that degrade intracellular proteins SHP2 and KRAS by recruiting them to ubiquitin-conjugating enzymes. This approach offers a new strategy for targeted protein degradation.
Area of Science:
- Biochemistry
- Molecular Biology
- Drug Discovery
Background:
- Targeted protein degradation is a therapeutic strategy.
- Biological Proteolysis-Targeting Chimeras (bioPROTACs) utilize E3 ligases for protein degradation.
- E2 ubiquitin-conjugating enzymes are crucial in the ubiquitination pathway.
Purpose of the Study:
- To engineer novel protein degraders by recruiting target proteins to E2 enzymes.
- To develop E2 bioPROTACs for the degradation of SHP2 and KRAS.
- To compare E2-based and VHL-based bioPROTACs using proteomics.
Main Methods:
- Rational design and screening of E2 bioPROTACs.
- Fusion of target-binding domains to human E2 enzymes.
- Global proteomics to analyze degradation effects.
- Protein display libraries for degrader discovery.
Main Results:
- Developed E2 bioPROTACs that induce degradation of SHP2 and KRAS.
- Characterized target-specific and global effects of E2 vs. VHL bioPROTACs.
- Identified a weak-affinity degrader via protein display that suppresses SHP2 signaling.
Conclusions:
- E2 bioPROTACs are a viable strategy for targeted protein degradation.
- This approach offers an alternative to E3 ligase-based systems.
- Protein display libraries can identify effective bioPROTACs with lower affinity.
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