Scout-MRM: Multiplexed Targeted Mass Spectrometry-Based Assay without Retention Time Scheduling Exemplified by Dickeya dadantii Proteomic Analysis during Plant Infection

Targeted mass spectrometry of a surrogate peptide panel is a powerful method to study the dynamics of protein networks, but chromatographic time scheduling remains a major limitation for dissemination and implementation of robust and large multiplexed assays. We unveil a Multiple Reaction Monitoring...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2017-02, Vol.89 (3), p.1421-1426
Main Authors: Rougemont, Blandine, Bontemps Gallo, Sébastien, Ayciriex, Sophie, Carrière, Romain, Hondermarck, Hubert, Lacroix, Jean Marie, Le Blanc, J. C. Yves, Lemoine, Jérôme
Format: Article
Language:English
Subjects:
Analytical chemistry
Chemical Sciences
Chromatography, High Pressure Liquid
Cichorium intybus - metabolism
Cichorium intybus - microbiology
Enterobacteriaceae - pathogenicity
Mass Spectrometry
Peptides - analysis
Plant Leaves - metabolism
Plant Leaves - microbiology
Proteomics - methods
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Summary:Targeted mass spectrometry of a surrogate peptide panel is a powerful method to study the dynamics of protein networks, but chromatographic time scheduling remains a major limitation for dissemination and implementation of robust and large multiplexed assays. We unveil a Multiple Reaction Monitoring method (Scout-MRM) where the use of spiked scout peptides triggers complex transition lists, regardless of the retention time of targeted surrogate peptides. The interest of Scout-MRM method regarding the retention time independency, multiplexing capability, reproducibility, and putative interest in facilitating method transfer was illustrated by a 782-peptide-plex relative assay targeting 445 proteins of the phytopathogen Dickeya dadantii during plant infection.
ISSN:0003-2700
1520-6882
DOI:10.1021/acs.analchem.6b03201