Top-down deep sequencing of Ubiquitin using two-dimensional mass spectrometry.
Fiche publication
Date publication
mai 2018
Journal
Analytical chemistry
Auteurs
Membres identifiés du Cancéropôle Est :
Dr DELSUC Marc-André
Tous les auteurs :
Floris F, Chiron L, Lynch A, Barrow MP, Delsuc MA, O'Connor PB
Lien Pubmed
Résumé
Two-dimensional Fourier Transform Ion Cyclotron Resonance Mass Spectrometry (2D FT-ICR MS or 2D MS) allows data independent fragmentation of all ions in a sample, and correlation of fragment ions to their precursors without isolation prior to fragmentation. Developments in computer capabilities and implementations in FTMS over of the last decade have allowed the technique to become a useful analytical tool for bottom-up proteomics (BUP), and more recently in top-down protein analysis (TDP). In this work, a new method of TDP is developed using two-dimensional mass spectrometry, called MS/2D FT-ICR MS or MS/2DMS. In MS/2DMS an entire protein is initially fragmented in a hexapole collision cell. The primary fragments are then sent to the ICR-cell, where 2D MS is performed with infrared multiphoton dissociation (IRMPD) or electron-capture dissociation (ECD). The resulting 2D mass spectrum retains information equivalent to a set of TDP MS3 experiments on the selected protein. MS/2DMS, and a related technique called MS/MS/2DMS, are used in this work to achieve 97% cleavage coverage and assignment for ubiquitin, noting several unique features which aid fragment identification.
Référence
Anal. Chem.. 2018 May 24;: