Post-acquisition ETD spectral processing for increased peptide identifications

J Am Soc Mass Spectrom. 2009 Aug;20(8):1435-40. doi: 10.1016/j.jasms.2009.03.006. Epub 2009 Mar 14.

Abstract

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(*)-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(*)-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 approximately 1% FDR increased from 4611 to 5931 upon spectral pre-processing--an increase of approximately 28.6%. We outline the most effective pre-processing methods, and provide free software containing these algorithms.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Algorithms*
  • Mass Spectrometry / methods*
  • Peptide Mapping / methods*
  • Peptides / chemistry*
  • Software*

Substances

  • Peptides