Systematic characterization of high mass accuracy influence on false discovery and probability scoring in peptide mass fingerprinting

Eric D. Dodds, Brian H. Clowers, Paul J Hagerman, Carlito B Lebrilla

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Whereas the bearing of mass measurement error on protein identification is sometimes underestimated, uncertainty in observed peptide masses unavoidably translates to ambiguity in subsequent protein identifications. Although ongoing instrumental advances continue to make high accuracy mass spectrometry (MS) increasingly accessible, many proteomics experiments are still conducted with rather large mass error tolerances. In addition, the ranking schemes of most protein identification algorithms do not include a meaningful incorporation of mass measurement error. This article provides a critical evaluation of mass error tolerance as it pertains to false positive peptide and protein associations resulting from peptide mass fingerprint (PMF) database searching. High accuracy, high resolution PMFs of several model proteins were obtained using matrix-assisted laser desorption/ionization Fourier transform ion cyclotron resonance mass spectrometry (MALDI-FTICR-MS). Varying levels of mass accuracy were simulated by systematically modulating the mass error tolerance of the PMF query and monitoring the effect on figures of merit indicating the PMF quality. Importantly, the benefits of decreased mass error tolerance are not manifest in Mowse scores when operating at tolerances in the low parts-per-million range but become apparent with the consideration of additional metrics that are often overlooked. Furthermore, the outcomes of these experiments support the concept that false discovery is closely tied to mass measurement error in PMF analysis. Clear establishment of this relation demonstrates the need for mass error-aware protein identification routines and argues for a more prominent contribution of high accuracy mass measurement to proteomic science.

Original languageEnglish (US)
Pages (from-to)156-166
Number of pages11
JournalAnalytical Biochemistry
Volume372
Issue number2
DOIs
StatePublished - Jan 15 2008

Fingerprint

Peptide Mapping
Peptides
Measurement errors
Proteins
Proteomics
Mass spectrometry
Mass Spectrometry
Bearings (structural)
Cyclotrons
Cyclotron resonance
Fourier Analysis
Uncertainty
Ionization
Desorption
Fourier transforms
Lasers
Experiments
Databases
Ions
Monitoring

Keywords

  • False positive
  • Fourier transform ion cyclotron resonance mass spectrometry
  • Mass measurement accuracy
  • Matrix-assisted laser desorption/ionization
  • Peptide mass fingerprinting

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Systematic characterization of high mass accuracy influence on false discovery and probability scoring in peptide mass fingerprinting. / Dodds, Eric D.; Clowers, Brian H.; Hagerman, Paul J; Lebrilla, Carlito B.

In: Analytical Biochemistry, Vol. 372, No. 2, 15.01.2008, p. 156-166.

Research output: Contribution to journalArticle

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