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Article Dans Une Revue Bioinformatics Année : 2010

Prophossi: automating expert validation of phosphopeptide–spectrum matches from tandem mass spectrometry

David M.A. Martin
  • Fonction : Auteur
Isabelle R.E. Nett
  • Fonction : Auteur
Michael A.J. Ferguson
  • Fonction : Auteur
David M A Martin
  • Fonction : Auteur
  • PersonId : 1083399
Isabelle R E Nett
  • Fonction : Auteur
Jonathan D Barber
  • Fonction : Auteur
Nicholas A Morrice
  • Fonction : Auteur
Michael a J Ferguson
  • Fonction : Auteur

Résumé

Motivation: Complex patterns of protein phosphorylation mediate many cellular processes. Tandem mass spectrometry (MS/MS) is a powerful tool for identifying these post-translational modifications. In high-throughput experiments, mass spectrometry database search engines, such as MASCOT provide a ranked list of peptide identifications based on hundreds of thousands of MS/MS spectra obtained in a mass spectrometry experiment. These search results are not in themselves sufficient for confident assignment of phosphorylation sites as identification of characteristic mass differences requires time-consuming manual assessment of the spectra by an experienced analyst. The time required for manual assessment has previously rendered high-throughput confident assignment of phosphorylation sites challenging. Results: We have developed a knowledge base of criteria, which replicate expert assessment, allowing more than half of cases to be automatically validated and site assignments verified with a high degree of confidence. This was assessed by comparing automated spectral interpretation with careful manual examination of the assignments for 501 peptides above the 1% false discovery rate (FDR) threshold corresponding to 259 putative phosphorylation sites in 74 proteins of the Trypanosoma brucei proteome. Despite this stringent approach, we are able to validate 80 of the 91 phosphorylation sites (88%) positively identified by manual examination of the spectra used for the MASCOT searches with a FDR < 15%. Conclusions: High-throughput computational analysis can provide a viable second stage validation of primary mass spectrometry database search results. Such validation gives rapid access to a systems level overview of protein phosphorylation in the experiment under investigation.
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Dates et versions

hal-03030170 , version 1 (29-11-2020)

Identifiants

Citer

David M.A. Martin, Isabelle R.E. Nett, Michael A.J. Ferguson, David M A Martin, Isabelle R E Nett, et al.. Prophossi: automating expert validation of phosphopeptide–spectrum matches from tandem mass spectrometry. Bioinformatics, 2010, 26 (17), pp.2153 - 2159. ⟨10.1093/bioinformatics/btq341⟩. ⟨hal-03030170⟩
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