Journal
JOURNAL OF THE AMERICAN SOCIETY FOR MASS SPECTROMETRY
Volume 20, Issue 8, Pages 1435-1440Publisher
SPRINGER
DOI: 10.1016/j.jasms.2009.03.006
Keywords
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Funding
- NIH [5T32GM08349]
- University of Wisconsin
- Beckman Foundation
- Eli Lilly
- National Institutes of Health [IR01GM080148]
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Tandem mass spectra (MS/MS) produced using electron transfer dissociation (ETD) differ from those derived from collision-activated dissociation (CAD) in several important ways. Foremost, the predominant fragment ion series are different: c- and z(center dot)-type ions are favored in ETD spectra while b- and y-type ions comprise the bulk of the fragments in CAD spectra. Additionally, ETD spectra possess charge-reduced precursors and unique neutral losses. Most database search algorithms were designed to analyze CAD spectra, and have only recently been adapted to accommodate c- and z(center dot)-type ions; therefore, inclusion of these additional spectral features can hinder identification, leading to lower confidence scores and decreased sensitivity. Because of this, it is important to pre-process spectral data before submission to a database search to remove those features that cause complications. Here, we demonstrate the effects of removing these features on the number Of unique peptide identifications at a 1% false discovery rate (FDR) using the open mass spectrometry search algorithm (OMSSA). When analyzing two biologic replicates of a yeast protein extract in three total analyses, the number of unique identifications with a similar to 1% FDR increased from 4611 to 5931 upon spectral preprocessing-an increase of similar to 28.6%. We outline the most effective pre-processing methods, and provide free software containing these algorithms. (J Am Soc Mass Spectrom 2009, 20, 1435-1440) (C) 2009 American Society for Mass Spectrometry
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