Mass Spectrometric Proteome Analysis of p53-dependent Neuronal Death

Proteomic analysis of p53-mediated neuronal cell death pathways. Advances in genomic analysis, including improvements in DNA sequencing, bioinformatics and the routine application of microarray technology to characterize gene expression profiles, have set the stage for understanding how genes are organized and regulated. However, the genetic blueprint cannot always predict which proteins are expressed, where they are localized in a cell and in what quantity and form they are present. Proteomics is a field of study that addresses these important and varied issues. Rapid innovations in the core technologies required to characterize proteins on a global scale are poised to bring about a comprehensive understanding of how dynamic changes in protein expression and function affect complex signaling and regulatory networks. We are currently applying a novel isotope coded affinity tag strategy in conjunction with high-resolution capillary liquid chromatography and mass spectrometry to characterize changes in the proteome of cultured mouse cortical neurons undergoing p53-dependent cell death after DNA damage. Thus far, the analysis has revealed the activation of opposing pro-apoptotic and anti-apoptotic processes after DNA damage in neurons. These results provide a remarkable overview of changes in functionally related cohorts of proteins during the process of cell death.