Halobacterium salinarum NRC-1 peptideAtlas: Toward strategies for targeted proteomics and improved proteome coverage

T. Van Phu, Amy K. Schmid, Nichole L. King, Amardeep Kaur, Min Pan, Kenia Whitehead, Tie Koide, Marc T. Facciotti, Young Ah Goo, Eric W. Deutsch, David J. Reiss, Parag Mallick, Nitin S. Baliga

Research output: Contribution to journalArticlepeer-review

31 Scopus citations

Abstract

The relatively small numbers of proteins and fewer possible post-translational modifications in microbes provide a unique opportunity to comprehensively characterize their dynamic proteomes. We have constructed a PeptideAtlas (PA) covering 62.7% of the predicted proteome of the extremely halophilic archaeon Halobacterium salinarum NRC-1 by compiling approximately 636 000 tandem mass spectra from 497 mass spectrometry runs in 88 experiments. Analysis of the PA with respect to biophysical properties of constituent peptides, functional properties of parent proteins of detected peptides, and performance of different mass spectrometry approaches has highlighted plausible strategies for improving proteome coverage and selecting signature peptides for targeted proteomics. Notably, discovery of a significant correlation between absolute abundances of mRNAs and proteins has helped identify low abundance of proteins as the major limitation in peptide detection. Furthermore, we have discovered that iTRAQ labeling for quantitative proteomic analysis introduces a significant bias in peptide detection by mass spectrometry. Therefore, despite identifying at least one proteotypic peptide for almost all proteins in the PA, a context-dependent selection of proteotypic peptides appears to be the most effective approach for targeted proteomics.

Original languageEnglish (US)
Pages (from-to)3755-3764
Number of pages10
JournalJournal of Proteome Research
Volume7
Issue number9
DOIs
StatePublished - Sep 2008

Keywords

  • Archaea
  • Bioinformatics
  • Halobacterium
  • Peptideatlas
  • Proteomics
  • iTRAQ

ASJC Scopus subject areas

  • Biochemistry
  • Chemistry(all)

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