Phosphorylation analysis of primary human T lymphocytes using sequential IMAC and titanium oxide enrichment

J Proteome Res. 2008 Dec;7(12):5167-76. doi: 10.1021/pr800500r.


T lymphocytes mediate cellular and humoral defense against foreign bodies or autoantigens. An understanding of T-cell information processing furthers studies of the immunological response. We describe a large-scale phosphorylation analysis of primary T cells using a multidimensional separation strategy, involving preparative SDS-PAGE for prefractionation, in-gel digestion and sequential phosphopeptide enrichment using IMAC and TiO2. A total of 281 phosphorylation sites (197 of high confidence, Ascore > 15), mapping to 204 human gene sequences, were identified by LC-MS(n) analysis in an LTQ linear ion trap. Subsequently, we created the LymPHOS database (, which links mass spectrometric peptide information to phosphorylation sites and phosphoprotein sequences.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Motifs
  • Amino Acid Sequence
  • Chromatography, Liquid / methods
  • Computational Biology / methods*
  • Humans
  • Immune System
  • Mass Spectrometry / methods
  • Molecular Sequence Data
  • Peptides / chemistry
  • Phosphopeptides / chemistry
  • Phosphorylation
  • Proteomics / methods*
  • T-Lymphocytes / metabolism*
  • Titanium / chemistry*
  • Tyrosine / chemistry


  • Peptides
  • Phosphopeptides
  • titanium dioxide
  • Tyrosine
  • Titanium