A practical recipe to survey phosphoproteomes

Abstract

The field of cellular signaling is fueled by the discovery of novel protein phosphorylation events. Phosphoproteomics focuses on the large-scale identification and characterization of serine, threonine, and tyrosine phosphorylation of proteins. Phosphopeptide enrichment followed by mass spectrometry has emerged as the most powerful technique for unbiased, discovery-driven analysis by offering high sensitivity, resolution, and speed. Methods for mass spectrometry-based phosphoproteomics analysis have improved substantially over the last decade, making the discipline more approachable to the broader scientific community. Herein we describe the status of the field of phosphoproteomics and provide a robust workflow covering the major aspects of large-scale phosphorylation analysis from phosphopeptide enrichment via IMAC to data analysis.