Extensive analysis of the cytoplasmic proteome of human erythrocytes using the peptide ligand library technology and advanced mass spectrometry

Florence Roux-Dalvai, Anne Gonzalez de Peredo, Carolina Simó, Luc Guerrier, David Bouyssié, Alberto Zanella, Attilio Citterio, Odile Burlet-Schiltz, Egisto Boschetti, Pier Giorgio Righetti, Bernard Monsarrat

Research output: Contribution to journalArticle

Abstract

The erythrocyte cytoplasmic proteome is composed of 98% hemoglobin; the remaining 2% is largely unexplored. Here we used a combinatorial library of hexameric peptides as a capturing agent to lower the signal of hemoglobin and amplify the signal of low to very low abundance proteins in the cytoplasm of human red blood cells (RBCs). Two types of hexapeptide library beads have been adopted: amino-terminal hexapeptide beads and beads in which the peptides have been further derivatized by carboxylation. The amplification of the signal of low abundance and suppression of the signal of high abundance species were fully demonstrated by two-dimensional gel maps and nano-LC-MSMS analysis. The effect of this new methodology on quantitative information also was explored. Moreover using this approach on an LTQ-Orbitrap mass spectrometer, we could identify with high confidence as many as 1578 proteins in the cytoplasmic fraction of a highly purified preparation of RBCs, allowing a deep exploration of the classical RBC pathways as well as the identification of unexpected minor proteins. In addition, we were able to detect the presence of eight different hemoglobin chains including embryonic and newly discovered globin chains. Thus, this extensive study provides a huge data set of proteins that are present in the RBC cytoplasm that may help to better understand the biology of this simplified cell and may open the way to further studies on blood pathologies using targeted approaches.

Original languageEnglish
Pages (from-to)2254-2269
Number of pages16
JournalMolecular and Cellular Proteomics
Volume7
Issue number11
DOIs
Publication statusPublished - Nov 2008

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Analytical Chemistry

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  • Cite this

    Roux-Dalvai, F., de Peredo, A. G., Simó, C., Guerrier, L., Bouyssié, D., Zanella, A., Citterio, A., Burlet-Schiltz, O., Boschetti, E., Righetti, P. G., & Monsarrat, B. (2008). Extensive analysis of the cytoplasmic proteome of human erythrocytes using the peptide ligand library technology and advanced mass spectrometry. Molecular and Cellular Proteomics, 7(11), 2254-2269. https://doi.org/10.1074/mcp.M800037-MCP200