Quantitative label-free phosphoproteomics strategy for multifaceted experimental designs.

ABSTRACT: Protein phosphorylation is a critical regulator of signaling in nearly all eukaryotic cellular pathways and dysregulated phosphorylation has been implicated in an array of diseases. The majority of MS-based quantitative phosphorylation studies are currently performed from transformed cell lines because of the ability to generate large amounts of starting material with incorporated isotopically labeled amino acids during cell culture. Here we describe a general label-free quantitative phosphoproteomic strategy capable of directly analyzing relatively small amounts of virtually any biological matrix, including human tissue and biological fluids. The strategy utilizes a TiO(2) enrichment protocol in which the selectivity and recovery of phosphopeptides were optimized by assessing a twenty-poi

SUBMITTER: Soderblom EJ 

PROVIDER: S-EPMC3093925 | biostudies-literature | 2011 May

REPOSITORIES: biostudies-literature