Proteome analysis of low-abundance proteins using multidimensional chromatography and isotope-coded affinity tags

Steven P. Gygi, Beate Rist, Timothy J. Griffin, Jimmy Eng, Ruedi Aebersold

Research output: Contribution to journalArticle

302 Scopus citations

Abstract

The effectiveness of proteome-wide protein identification and quantitative expression profiling is dependent on the ability of the analytical methodologies employed to routinely obtain information on low-abundance proteins, as these are frequently of great biological importance. Two-dimensional gel electrophoresis, the traditional method for proteome analysis, has proven to be biased toward highly expressed proteins. Recently, two-dimensional chromatography of the complex peptide mixtures generated by the digestion of unseparated protein samples has been introduced for the identification of their components, and isotope-coded affinity tags (ICAT) have been introduced to allow for accurate quantification of the components of protein mixtures by mass spectrometry. Here, we demonstrate that the combination of isotope coded affinity protein tags and multidimensional chromatography/mass spectrometry of tryptic peptide mixtures is capable of detecting and quantifying proteins of low abundance in complex samples.

Original languageEnglish (US)
Pages (from-to)47-54
Number of pages8
JournalJournal of Proteome Research
Volume1
Issue number1
DOIs
StatePublished - Jan 2002

Keywords

  • Functional genomics
  • Gene expression
  • Isotope-coded affinity tags
  • Mass spectrometry
  • Protein profiling
  • Proteomics

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