Fluorescent Biosensor for Detection of the R248Q Aggregation-Prone Mutant of p53

Area of Science:

• Molecular Biology
• Biochemistry
• Cancer Research

Background:

• The p53 tumor suppressor protein is frequently inactivated by missense mutations in human cancers.
• Mutations in the DNA-binding domain can cause conformational changes and amyloid-like aggregation.
• Detecting specific p53 mutants is crucial for understanding cancer development and progression.

Purpose of the Study:

• To engineer and characterize a fluorescent biosensor for the p53 R248Q mutant.
• To validate the biosensor's ability to detect the mutant protein in vitro and in living cells.
• To assess the biosensor's specificity for the R248Q mutant in different lung cancer cell lines.

Main Methods:

• Conjugation of an environmentally sensitive probe to a p53 peptide fragment.
• In vitro characterization of the fluorescent biosensor.
• Fluorescence microscopy for cellular detection after facilitated delivery.

Main Results:

• The developed biosensor successfully reports on the p53 R248Q mutant in vitro.
• The biosensor was effectively delivered and visualized in cultured cells.
• Preferential detection of the p53 R248Q mutant in PC9 lung cancer cells compared to other cell lines.

Conclusions:

• A novel fluorescent biosensor can specifically detect the p53 R248Q mutant.
• This biosensor shows promise for distinguishing cancer cells with specific p53 mutations.
• The tool aids in the study of p53 mutant behavior in cancer contexts.