Abstract
Protein phosphorylation is the major molecular mechanisms in the regulation of cellular functions and dynamically coordinate their signaling networks. Aberrant regulation of protein phosphorylation has been linked to a wide variety of human diseases, including cancer, immune abnormalities, and diabetes. With recent advances in liquid chromatography coupled to tandem mass spectrometry (LC-MS), phosphoproteome analysis has become a major area in biomedical research. Several chemoaffinity-based methods for enrichment of phosphopeptides have been developed, and in these methods, nucleic acids in the cells interfere with the binding of the phosphopeptides to affinity beads, thereby reducing the efficiency of enrichment. Here, we introduce a chemoaffinity-based method for enrichment of phosphopeptides in combination with protein extracts excluded of nucleic acids to improve the efficient enrichment of phosphopeptides.
