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Carbohydrate and Glycoconjugate Analysis: Studies of Protein N-Glycosylation and Oligosaccharide Isomer Discrimination by Mass Spectrometry and Ion Mobility
Abstract
Carbohydrates and glycoconjugates play key roles in cellular processes, including signaling, immune response, and influencing protein stability and function. Advances in mass spectrometry (MS)-based methods for the investigation of glycosylation and oligosaccharide structure are being made, however due to the level of structural diversity and complexity of these biomolecules detailed characterization remains a challenge. This dissertation can be generally grouped into two sections; N-glycopeptide analysis and carbohydrate analysis. The first four chapters focus on N-glycopeptide analysis by MS. Chapter 1 introduces the relevance and importance of glycoproteins as potential biomarkers for disease. Chapter 2 reviews a number of separations and MS-based methods for the analysis of glycopeptides. Chapter 3 describes a study of sialylated N-glycopeptides by a single tandem mass spectrometry (MS/MS) method, energy-resolved collision-induced dissociation (ER-CID). Chapter 4 demonstrates the application of electron transfer dissociation (ETD) and CID, in an ion mobility (IM)-resolved fashion to produce complementary fragmentation spectra in a single experiment. The next four chapters focus on the structural analysis of free carbohydrate isomers. Chapter 5 provides an overview of MS-based methods for the analysis of oligosaccharide isomers, including the use of IM and MS/MS, as standalone methods or when used in combination. Chapter 6 focuses on investigating the role that slight structural variations can play in the fragmentation pathways that the isomers may undergo. Chapter 7 describes the influence that transition metal cation adduction may play in the isomers’ unique fragmentation processes. Chapter 8 takes advantage of the ability to produce unique fragmentation spectra for two isomers and extends this ability to the ability to distinguish isomers in a mixture using IM. Overall, this dissertation includes a number of approaches using MS/MS, and occasionally IM, to fully characterize N-glycopeptides and allow for discrimination of oligosaccharide isomers.
Subject Area
Analytical chemistry
Recommended Citation
Schumacher, Katherine N, "Carbohydrate and Glycoconjugate Analysis: Studies of Protein N-Glycosylation and Oligosaccharide Isomer Discrimination by Mass Spectrometry and Ion Mobility" (2018). ETD collection for University of Nebraska-Lincoln. AAI10793426.
https://digitalcommons.unl.edu/dissertations/AAI10793426