Chemistry & Biochemistry Faculty Research and Publications
Effect of pH on In-Electrospray Hydrogen/Deuterium Exchange of Carbohydrates and Peptides
Document Type
Article
Publication Title
Journal of the American Society for Mass Spectrometry
Publication Date
Winter 2-7-2024
Volume
35
Issue
3
First Page
441
Last Page
448
Abstract
Carbohydrates are critical for cellular functions as well as an important class of metabolites. Characterizingcarbohydrate structures is a difficult analytical challenge due to the presence of isomers. In-electrospray hydrogen/deuteriumexchange mass spectrometry (in-ESI HDX-MS) is a method of HDX that samples the solvated structure of carbohydrates during theESI process and requires little to no instrument modification. Traditionally, solution-phase HDX is utilized with proteins to sampleconformational differences, and pH is a critical parameter to monitor and control due to the presence of both acid- and base-catalyzed mechanisms of exchange. For In-ESI HDX, the pH surrounding the analyte changes before and during labeling, which hasthe potential to affect the rate of labeling for analytes. Herein, we alter the pH of spray solutions containing model carbohydrates andpeptides, perform in-ESI HDX-MS, and characterize the deuterium uptake trends. Varying pH results in altered D uptake, thoughthe overall trends differ from the expected bulk-solution trends due to the electrospray process. These findings show the utility ofvarying pH prior to in-ESI HDX-MS for establishing different extents of HDX as well as distinguishing labile functional groups thatare present in different analytes.
Copyright held by
American Society for Mass Spectrometry
Recommended Citation
Effect of pH on In-Electrospray Hydrogen/Deuterium Exchange of Carbohydrates and Peptides Jacob B. Hatvany, et. al. Journal of the American Society for Mass Spectrometry 2024 35 (3), 441-448 DOI: 10.1021/jasms.3c00341
