Developing quantitative detection method for posttranslational modifications of p53 in colon cancer cell

Abstract

The tumor suppressor p53 plays a critical role in the response to cellular stress including DNA damage, hypoxia, oncogene activation and ribosomal stress. The p53 protein, which is transcriptionally modified by posttranslational modifications (PTMs), regulates hundreds of genes that control cell cycle arrest, apoptosis and senescence. Our study’s aim is to develop a quantitative detection method for PTMs of p53 in colon cancer cell using capillary electrophoresis coupled with laser-induced fluorescence (CE-LIF). We developed a quantitative CE-LIF method for the analysis of phosphorylated peptides of p53. We have analyzed 5 different 5-Carboxyfluorescein (5-FAM) labelled p53 peptides which was phosphorylated from different parts. Consequently, we have developed a sensitive, reproducible, cost effective and quantitative method for analysis of PTMs in p53. This methods can be further used in the detection of p53 in the cancer cells according to the LOD and LOQ values of validation results.