Related Experiment Video
Updated: Sep 26, 2025
10:55
Purification of Ubiquitinated p53 Proteins from Mammalian Cells
Published on: March 21, 2022
2.3K
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
Summary
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.
Related Concept Videos
01:13The Proteasome
1.2K
Eukaryotic cells can degrade proteins through several pathways. One of the most important among these is the ubiquitin-proteasome pathway. It helps the cell eliminate the misfolded, damaged, or unwarranted cytoplasmic proteins in a highly specific manner.
In this pathway, the target proteins are first tagged with small proteins called ubiquitin. This involves participation of a series of enzymes including— E1 (ubiquitin-activating enzyme), E2 (ubiquitin-conjugating enzyme), and E3...
In this pathway, the target proteins are first tagged with small proteins called ubiquitin. This involves participation of a series of enzymes including— E1 (ubiquitin-activating enzyme), E2 (ubiquitin-conjugating enzyme), and E3...
1.2K
02:58Regulated Protein Degradation
7.7K
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.
Protein degradation plays two important roles in the cells. It helps to protect cells from misfolded or damaged proteins before they lead to a...
Protein degradation plays two important roles in the cells. It helps to protect cells from misfolded or damaged proteins before they lead to a...
7.7K
02:04Covalently Linked Protein Regulators
7.5K
Proteins can undergo many types of post-translational modifications, often in response to changes in their environment. These modifications play an important role in the function and stability of these proteins. Covalently linked molecules include functional groups, such as methyl, acetyl, and phosphate groups, and also small proteins, such as ubiquitin. There are around 200 different types of covalent regulators that have been identified.
These groups modify specific amino acids in a protein....
These groups modify specific amino acids in a protein....
7.5K
01:17The Proteasome Structure
1.0K
The ubiquitin-proteasome pathway is a well-known mechanism utilized by eukaryotic cells to remove cytoplasmic proteins that are misfolded, damaged, or no longer needed. In this pathway, the protein that needs to be eliminated undergoes a process called ubiquitination, where a chain of ubiquitin molecules is attached to the 48th lysine residue of the target protein. This ubiquitin modification helps the proteasome distinguish between a target protein and a healthy protein.
The proteasome is an...
The proteasome is an...
1.0K
01:32Export of Misfolded Proteins out of the ER
4.0K
After folding, the ER assesses the quality of secretory and membrane proteins. The correctly folded proteins are cleared by the calnexin cycle for transport to their final destination, while misfolded proteins are held back in the ER lumen. The ER chaperones attempt to unfold and refold the misfolded proteins but sometimes fail to achieve the correct native conformation. Such terminally misfolded proteins are then exported to the cytosol by ER-associated degradation or ERAD pathway for...
4.0K
01:57Protein Complexes with Interchangeable Parts
2.6K
Groups of proteins may form a complex where each protein in this complex has a different role in the overall execution of the complex’s function. Often some of the proteins in the complex can be replaced by a closely related variant to give a complex that contains many of the same components yet is functionally distinct.
The SCF ubiquitin ligase is a protein complex of five individual proteins. This complex attaches ubiquitin to other target proteins to mark them for degradation. In order...
The SCF ubiquitin ligase is a protein complex of five individual proteins. This complex attaches ubiquitin to other target proteins to mark them for degradation. In order...
2.6K