Fast alignment of mass spectra in large proteomics datasets, capturing dissimilarities arising from multiple complex modifications of peptides

Abstract

ABSTRACTBackgroundIn proteomics, the interpretation of mass spectra representing peptides carrying multiple complex modifications is still challenging, currently limited by the number of potential modifications considered in a single analysis and the need to know them in advance. Further developments must be done in the field to help the scientific community to discover new post-translational modifications that play an essential role in disease and to understand how chemical modifications carried by food proteins could impact our health.ResultsTo make progress on this issue, we implemented SpecGlobX (SpecGlob eXTended to eXperimental spectra), a standalone Java application that quickly determines the best spectral alignments of a (possibly very large) list of Peptide-to-Spectrum Matches (PSMs) provided by any open modification search method, or generated by the user. As input, SpecGlobX reads a file containing spectra in MGF or mzML format and a semicolon-delimited spreadsheet describing the PSMs. As output, SpecGlobX returns the best alignment for each PSM, splitting the mass difference between the spectrum and the peptide into one or more shifts while considering the possibility of non-aligned masses (a phenomenon resulting from many situations including neutral losses).SpecGlobX is fast, able to align one million PSMs in about 1.5 minutes on a standard desktop. Firstly, we remind the foundations of the algorithm and detail how we adapted SpecGlob (the method we previously developed following the same aim, but limited to the interpretation of perfect simulated spectra) to the interpretation of imperfect experimental spectra. Then, we highlight the interest of SpecGlobX as a complementary tool downstream to three open modification search methods on a large simulated spectra dataset. Finally, we show on a smaller dataset that SpecGlobX performs equally well on experimental and simulated spectra.ConclusionsSpecGlobX is helpful as a decision support tool, providing keys to interpret peptides carrying complex modifications still poorly considered by current open modification search software. Better alignment of PSMs enhances confidence in the identification of spectra provided by open modification search methods and should improve the interpretation rate of spectra.