New algorithm for 15N//14N quantitation with LC-ESI-MS using an LTQ-FT mass spectrometer

Victor P. Andreev, Lingyun Li, Tomas Rejtar, Qingbo Li, James G. Ferry, Barry L. Karger

Research output: Contribution to journalArticle

31 Scopus citations

Abstract

A new algorithm (QN) for the 15N/14N quantitation of relative protein abundances in complex proteomic samples is described. QN takes advantage of the high resolution, mass accuracy and throughput of the hybrid mass spectrometer LTQ-FT MS. Peptide quantitation is based on MS peak intensity (measured in the FT MS), while peptide identification is performed in the MS/MS mode (measured in the LTQ linear ion trap). Accuracy of the protein abundance is enhanced by a novel scoring procedure, allowing filtering of less reliable measurements of peptide abundances. The performance of QN is illustrated in the relative quantitative analysis of M. acetivorans C2A cultures grown with carbon monoxide vs methanol as substrate. Roughly 1000 proteins were quantitated with an average CV of 9% for the protein abundance ratios. QN performs quantitation without manual intervention, does not require high processing power, and generates files compatible with the Guidelines for Proteomic Data Publication.

Original languageEnglish (US)
Pages (from-to)2039-2045
Number of pages7
JournalJournal of Proteome Research
Volume5
Issue number8
DOIs
StatePublished - Aug 1 2006

Keywords

  • LC-MS
  • M. acetivorans
  • Metabolic labeling
  • Quantitative proteomics
  • Software

ASJC Scopus subject areas

  • Genetics
  • Biotechnology
  • Biochemistry

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    Andreev, V. P., Li, L., Rejtar, T., Li, Q., Ferry, J. G., & Karger, B. L. (2006). New algorithm for 15N//14N quantitation with LC-ESI-MS using an LTQ-FT mass spectrometer. Journal of Proteome Research, 5(8), 2039-2045. https://doi.org/10.1021/pr060105m