Benchmarking common quantification strategies for large-scale phosphoproteomics.

ABSTRACT: Comprehensive mass spectrometry (MS)-based proteomics is now feasible, but reproducible quantification remains challenging, especially for post-translational modifications such as phosphorylation. Here, we compare the most popular quantification techniques for global phosphoproteomics: label-free quantification (LFQ), stable isotope labeling by amino acids in cell culture (SILAC) and MS²- and MS³-measured tandem mass tags (TMT). In a mixed species comparison with fixed phosphopeptide ratios, we find LFQ and SILAC to be the most accurate techniques. MS²-based TMT yields the highest precision but lowest accuracy due to ratio compression, which MS³-based TMT can partly rescue. However, MS²-based TMT outperforms MS³-based TMT wh

SUBMITTER: Hogrebe A 

PROVIDER: S-EPMC5849679 | biostudies-literature | 2018 Mar

REPOSITORIES: biostudies-literature