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Yeast As a Chassis for Developing Functional Assays to Study Human P53
Published on: August 4, 2019
Promising Proteolysis-Targeting Chimera for Mutant p53-R175H
Xinzhe Zhuang1, Yidan Guo1, Xiaozi Sun1
1School of Life Sciences and Medicine, Shandong University of Technology, Zibo 255000, China.
Abstract:
The tumor suppressor protein p53 is among the most commonly mutated proteins across a variety of cancer types. Notably, the p53 R175H mutation ranks as one of the most prevalent hotspot mutations. Proteolysis-targeting chimeras (PROTACs) represent a class of bifunctional molecules capable of harnessing the cellular ubiquitin-proteasome pathway to facilitate targeted protein degradation. Despite the potential of PROTACs, limited research has been directed toward the degradation of the p53-R175H mutant protein. In this study, we developed a series of peptide-based PROTACs, leveraging known peptide ligands for both the p53-R175H mutation and the E3 ubiquitin ligase VHL. Our findings indicate that one of these peptide-based PROTACs is capable of directing the p53-R175H protein to the proteasome for degradation within a recombinant expression system. Moreover, by synthesizing a fusion peptide PROTAC molecule that incorporates a membrane-penetrating peptide, we have demonstrated its ability to traverse cellular membranes and subsequently reduce the levels of the p53-R175H mutant protein. Importantly, the degradation of p53-R175H was found to mitigate the cellular migration and invasion. In summary, our study introduces a novel class of protein degraders and establishes a foundational framework for the therapeutic management of cancers associated with p53 mutations.
Insights
Researchers developed novel peptide-based PROTACs to degrade the mutant p53 R175H protein, a common cancer mutation. These PROTACs effectively reduced tumor suppressor protein levels and mitigated cancer cell migration and invasion.
Area of Science:
- Oncology
- Molecular Biology
- Biochemistry
Background:
- The tumor suppressor protein p53 is frequently mutated in various cancers, with the R175H mutation being a prevalent hotspot.
- Proteolysis-targeting chimeras (PROTACs) are bifunctional molecules that induce targeted protein degradation via the ubiquitin-proteasome system.
- Targeted degradation of mutant p53, specifically the R175H variant, remains an underexplored therapeutic strategy.
Purpose of the Study:
- To design and synthesize novel peptide-based PROTACs targeting the p53 R175H mutant protein.
- To evaluate the efficacy of these PROTACs in degrading p53 R175H in vitro and in cell-based systems.
- To assess the functional consequences of p53 R175H degradation on cancer cell behavior.
Main Methods:
- Development of peptide-based PROTACs utilizing ligands for p53 R175H and the E3 ligase VHL.
- Testing PROTAC-mediated degradation in a recombinant expression system.
- Synthesis of a membrane-penetrating peptide PROTAC for cellular studies.
- Assessment of cellular migration and invasion assays.
Main Results:
- A novel peptide-based PROTAC successfully induced the degradation of p53 R175H in a recombinant system.
- A fusion peptide PROTAC demonstrated membrane permeability and reduced p53 R175H levels in cells.
- Degradation of p53 R175H led to a significant decrease in cancer cell migration and invasion.
Conclusions:
- This study introduces a new class of peptide-based PROTACs for targeted degradation of the p53 R175H mutant protein.
- The developed PROTACs show potential for therapeutic intervention in cancers harboring p53 mutations.
- This work provides a foundation for developing PROTAC-based therapies against p53-mutated cancers.
