The divide and conquer strategies for deep phosphoproteomics analysis

With the advance of mass spectrometry based proteomics techniques, thousands of phosphorylation sites can be identified from a single cell line or a single tissue sample by large scale global analysis. However, such large scale analysis is time consuming. And, due to the high complexity of the phosphoproteome sample, this approach is often unable to identify low abundance regulatory phosphorylation sites. Alternatively, the “divide and conquer” strategy by targeted analysis of a subset of phosphoproteome, i.e. subphosphoproteome, can improve the detection sensitivity of these phosphorylation sites. In this review, we summarized the analytical methods in this field. The enrichment approaches for the analysis of subphosphoproteomes generated by tyrosine kinases and basophilic kinases were introduced. The strategies for the analysis of the subphosphoproteomes composed by endogenous phosphopeptides, multiply phosphorylated peptides, etc., were also reviewed.