Journal
JOURNAL OF PROTEOME RESEARCH
Volume 9, Issue 11, Pages 5511-5516Publisher
AMER CHEMICAL SOC
DOI: 10.1021/pr1006813
Keywords
mass accuracy; HCD; identification
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Funding
- Bundesministerium fur Bildung und Forschung, Spitzencluster BioRN, Verbundprojekt Inkubator [BioRN-INE-TP01, BioRN-IND-TP02]
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Currently, scoring algorithms of many popular search engines for tandem mass spectrometry (MS/MS) data only partially utilize the information content of high mass accuracy MS/MS data. We have developed a new rescoring scheme, H-score, that employs high mass accuracy matching of all detected fragment ions to candidate peptide sequences in an abundance independent fashion. Peptides for which b or y ions are found for all or almost all backbone fragmentation sites are rewarded. For peptide hits generated by Mascot, rescoring proved to be particularly beneficial when applied on samples containing many different potential modifications. For a histone sample acquired on an Orbitrap Velos using HCD for peptide fragmentation, the H-score identified 24% more spectra at 0.01 false positive rate than Mascot scoring of spectra processed according to state-of-the-art methods and 61% better than Mascot scoring of unprocessed MS/MS spectra. For a low-abundance sample, where many weak spectra were detected, these numbers went up to 53 and 190%, respectively. When applied on a kinase-enriched sample containing only a few modifications, a smaller but still significant gain of 5% was observed.
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