Large-scale characterization of HeLa cell nuclear phosphoproteins Academic Article uri icon


MeSH Major

  • Nuclear Proteins
  • Phosphoproteins


  • Determining the site of a regulatory phosphorylation event is often essential for elucidating specific kinase-substrate relationships, providing a handle for understanding essential signaling pathways and ultimately allowing insights into numerous disease pathologies. Despite intense research efforts to elucidate mechanisms of protein phosphorylation regulation, efficient, large-scale identification and characterization of phosphorylation sites remains an unsolved problem. In this report we describe an application of existing technology for the isolation and identification of phosphorylation sites. By using a strategy based on strong cation exchange chromatography, phosphopeptides were enriched from the nuclear fraction of HeLa cell lysate. From 967 proteins, 2,002 phosphorylation sites were determined by tandem MS. This unprecedented large collection of sites permitted a detailed accounting of known and unknown kinase motifs and substrates.

publication date

  • August 17, 2004



  • Academic Article



  • eng

PubMed Central ID

  • PMC514446

Digital Object Identifier (DOI)

  • 10.1073/pnas.0404720101

PubMed ID

  • 15302935

Additional Document Info

start page

  • 12130

end page

  • 5


  • 101


  • 33