Enrichment strategy development for phosphoproteome analysis of saccharomyces cerevisiae

Abstract:

The reversible phosphorylation of proteins is central to regulating most aspects of cell function. Malfunction in this critical cellular process have been implicated to cause diseases such as diabetes, cancer, and Alzheimer’s. Recent advances in mass spectrometry have made it possible to study this important post translational modification on a proteome-wide scale. However, to be able to do so, enrichment of phosphorylated peptides is required. Pairwise comparison of individual steps in an enrichment procedure and simultaneous improvement of data analysis resulted in a protocol which allowed high confidence identification of 2,131 unique phosphorylated peptides from 1,026 proteins. Thereby not only establishing a working protocol for phosphopeptide enrichment in the Griffin Lab, but also generating the largest list of proteins phosphorylated under normal conditions in yeast to date.