Premium
Ultra‐high intra‐spectrum mass accuracy enables unambiguous identification of fragment reporter ions in isobaric multiplexed quantitative proteomics
Author(s) -
Pachl Fiona,
Fellenberg Kurt,
Wagner Corinna,
Kuster Bernhard
Publication year - 2012
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.201100622
Subject(s) - isobaric labeling , isobaric process , tandem mass tag , mass spectrometry , tandem mass spectrometry , mass spectrum , quantitative proteomics , fragmentation (computing) , chemistry , tandem , ion , proteomics , top down proteomics , analytical chemistry (journal) , protein mass spectrometry , chromatography , computer science , physics , materials science , biochemistry , organic chemistry , composite material , gene , thermodynamics , operating system
Isobaric tagging using reagents such as tandem mass tags ( TMT ) and isobaric tags for relative and absolute quantification (i TRAQ ) have become popular tools for mass spectrometry based quantitative proteomics. Because the peptide quantification information is collected in tandem mass spectra, the accuracy and precision of this method largely depend on the resolution with which precursor ions can be selected for the fragmentation and the specificity of the generated reporter ion. The latter can constitute an issue if near isobaric ion signals are present in such spectra because they may distort quantification results. We propose a simple remedy for this problem by identifying reporter ions via the accurate mass differences within a single tandem mass spectrum instead of applying fixed mass error tolerances for all tandem mass spectra. Our results show that this leads to unambiguous reporter ion identification and complete removal of interfering signals. This mode of data processing is easily implemented in software and offers advantages for protein quantification based on few peptides.