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Electron‐Activated Tandem Mass Spectrometry Analysis of Glycosaminoglycans
Author(s) -
Pepi Lauren E.,
Amster I. Jonathan
Publication year - 2021
Publication title -
current protocols
Language(s) - English
Resource type - Journals
ISSN - 2691-1299
DOI - 10.1002/cpz1.83
Subject(s) - glycosaminoglycan , mass spectrometry , tandem mass spectrometry , chemistry , tandem mass tag , tandem , chromatography , materials science , biochemistry , proteomics , quantitative proteomics , composite material , gene
Abstract Glycosaminoglycans (GAGs) are linear polysaccharides found in a variety of organisms. GAGs contribute to biochemical pathway regulation, cell signaling, and disease progression. GAG sequence information is imperative for determining structure‐function relationships. Recent advances in electron‐activation techniques paired with high‐resolution mass spectrometry allow for full sequencing of GAG structures. Electron detachment dissociation (EDD) and negative electron transfer dissociation (NETD) are two electron‐activation methods that have been utilized for GAG characterization. Both methods produce an abundance of informative glycosidic and cross‐ring fragment ions without producing a high degree of sulfate decomposition. Here, we provide detailed protocols for using EDD and NETD to sequence GAG chains. In addition to protocols directly involving performing these MS/MS methods, protocols include sample preparation, method development, internal calibration, and data analysis. © 2021 Wiley Periodicals LLC. This article was corrected on 27 July 2022. See the end of the full text for details. Basic Protocol 1 : Preparation of glycosaminoglycan samples Basic Protocol 2 : FTICR method development Basic Protocol 3 : Internal calibration with NaTFA Basic Protocol 4 : Electron Detachment Dissociation (EDD) of GAG samples Basic Protocol 5 : Negative electron transfer dissociation (NETD) of GAG samples Basic Protocol 6 : Analysis of MS/MS data