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

Blandine Rougemont (1) , Sebastien B Gallo (2) , Sophie Ayciriex (1) , Romain Carrière (1) , Hubert Hondermarck (3, 4) , Jean-Marie Lacroix (2) , J.C.Y. Leblanc (5) , Jérôme Lemoine (1)

1 ISA-ANABIO-MS - Analyse biomoléculaire par spectrométrie de masse - Biological Analysis by Mass Spectrometry
2 UGSF - Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576
3 Signalisation des facteurs de croissance dans le cancer du sein. Protéomique fonctionnelle
4 IBLI - Institut de biologie de Lille - IBL
5 SCIEX

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.

Mots clés: Scout-MRM; Proteomics; plant infection; Dickeya dadantii; Multiplexed Targeted Mass Spectrometry

Domaines: Chimie analytique

Blandine Rougemont, Sebastien B Gallo, Sophie Ayciriex, Romain Carrière, Hubert Hondermarck, et al.. Scout-MRM: Multiplexed Targeted Mass Spectrometry-Based Assay without Retention Time Scheduling Exemplified by Dickeya dadantii Proteomic Analysis during Plant Infection. Analytical Chemistry, 2017, 89 (3), pp.1421-1426 ⟨10.1021/acs.analchem.6b03201⟩. ⟨hal-01518912⟩