Combined use of Ion Mobility and Collision-Induced Dissociation to investigate the opening of disulfide bridges by Electron-Transfer Dissociation in peptides bearing two disulfide bonds
[en] Disulfide bonds are post-translational modifications (PTMs) often found in peptides and proteins. They increase their stability towards enzymatic degradations, and provide the structure and (consequently) the activity of such folded proteins. The characterization of disulfide patterns, i.e the cysteine connectivity, is crucial to achieve a global picture of the active conformation of the protein of interest. Electron Transfer Dissociation (ETD) constitutes a valuable tool to cleave the disulfide bonds in the gas phase, avoiding chemical reduction/alkylation in solution. To characterize the cysteine pairing, the present work proposes (i) to reduce by ETD one of the two disulfide bridges of model peptides , resulting in the opening of the cyclic structures, (ii) to separate the generated species by ion-mobility and, (iii) to characterize the species using CID. Results of this strategy applied to several peptides show different behaviors depending on the connectivity. The loss of SH• radical species, observed for all the peptides, confirms the cleavage of the disulfides during the ETD process.
Research center :
Giga-Systems Biology and Chemical Biology - ULiège CART - Centre Interfacultaire d'Analyse des Résidus en Traces - ULiège
Massonnet, Philippe ; Université de Liège > Département de chimie (sciences) > Laboratoire de spectrométrie de masse (L.S.M.)
Upert, Gregory; CEA Saclay > CEA/DSV/iBiTec-S/SIMOPRO Bat 152 91191 Gif sur Yvette > Toxins, Receptors and Channels
Smargiasso, Nicolas ; Université de Liège > Département de chimie (sciences) > Laboratoire de spectrométrie de masse (L.S.M.)
Gilles, Nicolas; CEA Saclay > CEA/DSV/iBiTec-S/SIMOPRO Bat 152, 91191 Gif sur Yvette > Toxins, Receptors and Channels
Quinton, Loïc ; Université de Liège > Département de chimie (sciences) > Chimie biologique
De Pauw, Edwin ; Université de Liège > Département de chimie (sciences) > Laboratoire de spectrométrie de masse (L.S.M.)
Language :
English
Title :
Combined use of Ion Mobility and Collision-Induced Dissociation to investigate the opening of disulfide bridges by Electron-Transfer Dissociation in peptides bearing two disulfide bonds
Publication date :
2015
Journal title :
Analytical Chemistry
ISSN :
0003-2700
eISSN :
1520-6882
Publisher :
American Chemical Society, Washington, United States - District of Columbia
FP7 - 278346 - VENOMICS - High-throughput peptidomics and transcriptomics of animal venoms for discovery of novel therapeutic peptides and innovative drug development
Name of the research project :
VENOMICS
Funders :
FRIA - Fonds pour la Formation à la Recherche dans l'Industrie et dans l'Agriculture [BE] CE - Commission Européenne [BE]
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