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Inhibition of p53 Mutant Peptide Aggregation In Vitro by Cationic Osmolyte Acetylcholine Chloride
Zhaolin Chen, Mathumai Kanapathipillai1
1University of Michigan- Dearborn, Department of Mechanical Engineering, 4901, Evergreen Road, Dearborn, MI 48128. United States.
Abstract:
Mutations of tumor suppressor protein p53 are present in almost about 50% of all cancers. It has been reported that the p53 mutations cause aggregation and subsequent loss of p53 function, leading to cancer progression. Here in this study we focus on the inhibitory effects of cationic osmolyte molecules acetylcholine chloride, and choline on an aggregation prone 10 amino acid p53 mutant peptide WRPILTIITL, and the corresponding wildtype peptide RRPILTIITL in vitro. The characterization tools used for this study include Thioflavin- T (ThT) induced fluorescence, transmission electron microscopy (TEM), congo red binding, turbidity, dynamic light scattering (DLS), and cell viability assays. The results show that acetylcholine chloride in micromolar concentrations significantly inhibit p53 mutant peptide aggregation in vitro, and could be promising candidate for p53 mutant/ misfolded protein aggregation inhibition.
Insights
Acetylcholine chloride effectively inhibits the aggregation of mutant p53 peptides in vitro. This finding suggests its potential as a therapeutic agent for cancers linked to p53 mutations and protein misfolding.
Area of Science:
- Biochemistry
- Molecular Biology
- Cancer Research
Background:
- Mutations in the tumor suppressor protein p53 occur in approximately 50% of human cancers.
- p53 mutations lead to protein aggregation, loss of function, and cancer progression.
Purpose of the Study:
- To investigate the inhibitory effects of cationic osmolytes acetylcholine chloride and choline on the aggregation of a p53 mutant peptide.
- To evaluate the potential of these molecules in preventing p53 mutant/misfolded protein aggregation.
Main Methods:
- In vitro study using a 10 amino acid p53 mutant peptide (WRPILTIITL) and its wildtype counterpart (RRPILTIITL).
- Characterization techniques included Thioflavin-T fluorescence, transmission electron microscopy (TEM), Congo red binding, turbidity, and dynamic light scattering (DLS).
- Cell viability assays were also performed.
Main Results:
- Acetylcholine chloride demonstrated significant inhibition of p53 mutant peptide aggregation at micromolar concentrations.
- Choline's effects were also assessed, though acetylcholine chloride showed more pronounced inhibition.
- The tested molecules did not show significant toxicity in cell viability assays at inhibitory concentrations.
Conclusions:
- Acetylcholine chloride is a promising candidate for inhibiting p53 mutant/misfolded protein aggregation.
- Targeting protein aggregation with small molecules like acetylcholine chloride offers a potential therapeutic strategy for p53-mutated cancers.
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