Electron-Activated Tandem Mass Spectrometry Analysis of Glycosaminoglycans.

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. 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.