1. Benefits of Iterative Searches of Large Databases to Interpret Large Human Gut Metaproteomic Data Sets
- Author
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Didier Chevret, Magali Berland, Sandra Plancade, Sylvie Huet, Mélisande Blein-Nicolas, Alain Guillot, Ariane Bassignani, Karine Clément, Catherine Juste, Olivier Langella, Salwa W. Rizkalla, Joël Doré, Chloé Moritz, Génétique Quantitative et Evolution - Le Moulon (Génétique Végétale) (GQE-Le Moulon), and AgroParisTech-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)
- Subjects
Proteomics ,0301 basic medicine ,Matching (statistics) ,Simple Modular Architecture Research Tool ,Computer science ,[SDV]Life Sciences [q-bio] ,computer.software_genre ,Biochemistry ,03 medical and health sciences ,Human health ,Human gut ,Tandem Mass Spectrometry ,Humans ,Database search engine ,Microbiome ,ComputingMilieux_MISCELLANEOUS ,030102 biochemistry & molecular biology ,Database ,Microbiota ,General Chemistry ,Gastrointestinal Microbiome ,030104 developmental biology ,Metagenomics ,Metaproteomics ,computer - Abstract
The gut microbiota are increasingly considered as a main partner of human health. Metaproteomics enables us to move from the functional potential revealed by metagenomics to the functions actually operating in the microbiome. However, metaproteome deciphering remains challenging. In particular, confident interpretation of a myriad of MS/MS spectra can only be pursued with smart database searches. Here, we compare the interpretation of MS/MS data sets from 48 individual human gut microbiomes using three interrogation strategies of the dedicated Integrated nonredundant Gene Catalog (IGC 9.9 million genes from 1267 individual fecal samples) together with the Homo sapiens database: the classical single-step interrogation strategy and two iterative strategies (in either two or three steps) aimed at preselecting a reduced-sized, more targeted search space for the final peptide spectrum matching. Both iterative searches outperformed the single-step classical search in terms of the number of peptides and protein clusters identified and the depth of taxonomic and functional knowledge, and this was the most convincing with the three-step approach. However, iterative searches do not help in reducing variability of repeated analyses, which is inherent to the traditional data-dependent acquisition mode, but this variability did not affect the hierarchical relationship between replicates and all other samples.
- Published
- 2021
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