Abstract
Analysis pipelines that assign peptides to shotgun proteomics mass spectra often discard identified spectra deemed irrelevant to the scientific hypothesis being tested. To improve statistical power, I propose that researchers remove irrelevant peptides from the database prior to searching rather than assigning these peptides to spectra and then discarding the matches.
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Acknowledgements
I thank G. Knudsen for providing access to the honeybee data set. This work was funded by US National Institutes of Health awards R01 GM096306 and P41 GM103533.
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Supplementary information
Supplementary texts
Supplementary Methods (PDF 365 kb)
Supplementary Data 1
Reanalyzed Plasmodium data. The zip file contains five files: two lists of tryptic peptides (Plasmodium and human) and three lists of peptide-spectrum matches (Plasmodium, human, and Plasmodium+human). (ZIP 11716 kb)
Supplementary Data 2
Reanalyzed SP-A data. The zip file contains five files: two lists of tryptic peptides (SP_A variants, human proteome) and three lists of peptide-spectrum matches (SP_A, human, SP_A+human). (ZIP 37844 kb)
Supplementary Data 3
Reanalyzed honeybee data. The zip file contains five files: two lists of tryptic peptides (virus and honeybee) and three lists of peptide-spectrum matches (virus, honeybee, virus+honeybee). (ZIP 46845 kb)
Supplementary Data 4
Source data for Figure 1. (ZIP 609 kb)
Supplementary Data 5
Source data for Figure 2. (ZIP 25 kb)
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Noble, W. Mass spectrometrists should search only for peptides they care about. Nat Methods 12, 605–608 (2015). https://doi.org/10.1038/nmeth.3450
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DOI: https://doi.org/10.1038/nmeth.3450
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