Doublet N-Terminal Oriented Proteomics for N-Terminomics and Proteolytic Processing Identification.
Fiche publication
Date publication
janvier 2017
Journal
Methods in molecular biology (Clifton, N.J.)
Auteurs
Membres identifiés du Cancéropôle Est :
Mme SCHAEFFER-REISS Christine, Dr CARAPITO Christine
Tous les auteurs :
Westermann B, Jacome ASV, Rompais M, Carapito C, Schaeffer-Reiss C
Lien Pubmed
Résumé
The study of the N-terminome and the precise identification of proteolytic processing events are key in biology. Dedicated methodologies have been developed as the comprehensive characterization of the N-terminome can hardly be achieved by standard proteomics methods. In this context, we have set up a trimethoxyphenyl phosphonium (TMPP) labeling approach that allows the characterization of both N-terminal and internal digestion peptides in a single experiment. This latter point is a major advantage of our strategy as most N-terminomics methods rely on the enrichment of N-terminal peptides and thus exclude internal peptides.We have implemented a double heavy/light TMPP labeling and an automated data validation workflow that make our doublet N-terminal oriented proteomics (dN-TOP) strategy efficient for high-throughput N-terminome analysis.
Mots clés
Chromatography, Liquid, methods, Isotope Labeling, Peptide Fragments, Proteolysis, Proteome, Proteomics, methods, Statistics as Topic, methods, Tandem Mass Spectrometry, methods, Workflow
Référence
Methods Mol. Biol.. 2017 ;1574:77-90