Increasing information from shotgun proteomic data by accounting for misassigned precursor ion masses

Proteomics. 2008 Jul;8(14):2791-7. doi: 10.1002/pmic.200800045.

Abstract

Although mass spectrometers are capable of providing high mass accuracy data, assignment of true monoisotopic precursor ion mass is complicated during data-dependent ion selection for LC-MS/MS analysis of complex mixtures. The complication arises when chromatographic peak widths for a given analyte exceed the time required to acquire a precursor ion mass spectrum. The result is that many measured monoisotopic masses are misassigned due to calculation from a single mass spectrum with poor ion statistics based on only a fraction of the total available ions for a given analyte. Such data in turn produces errors in automated database searches, where precursor m/z value is one search parameter. We propose here a postacquisition approach to correct misassigned monoisotopic m/z values that involves peak detection over the entire elution profile and correction of the precursor ion monoisotopic mass. As a result of using this approach to reprocess shotgun proteomic data we increased peptide sequence assignments by 10% while reducing the estimated false positive ratio from 1 to 0.2%. We also show that 4% of the salvaged identifications may be accounted for by correction of mixed tandem mass spectra resulting from fragmentation of multiple peptides simultaneously, a situation which we refer to as accidental CID.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural

MeSH terms

  • Bacterial Proteins / analysis
  • Bacterial Proteins / standards
  • Chromatography, Liquid / methods
  • Chromatography, Liquid / standards
  • Computational Biology / methods*
  • Computational Biology / standards
  • False Positive Reactions
  • Ions / analysis*
  • Ions / chemistry
  • Molecular Weight
  • Proteomics / methods*
  • Proteomics / standards
  • Pseudomonas aeruginosa / chemistry
  • Spectrometry, Mass, Electrospray Ionization / methods
  • Spectrometry, Mass, Electrospray Ionization / standards
  • Tandem Mass Spectrometry / methods
  • Tandem Mass Spectrometry / standards

Substances

  • Bacterial Proteins
  • Ions