Characterization of native protein complexes and protein isoform variation using size-fractionation-based quantitative proteomics.

ABSTRACT: Proteins form a diverse array of complexes that mediate cellular function and regulation. A largely unexplored feature of such protein complexes is the selective participation of specific protein isoforms and/or post-translationally modified forms. In this study, we combined native size-exclusion chromatography (SEC) with high-throughput proteomic analysis to characterize soluble protein complexes isolated from human osteosarcoma (U2OS) cells. Using this approach, we have identified over 71,500 peptides and 1,600 phosphosites, corresponding to over 8,000 proteins, distributed across 40 SEC fractions. This represents >50% of the predicted U2OS cell proteome, identified with a mean peptide sequence coverage of 27% per protein. Three biological replicates were performed, allowing statistical

SUBMITTER: Kirkwood KJ 

PROVIDER: S-EPMC3861729 | biostudies-literature | 2013 Dec

REPOSITORIES: biostudies-literature