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Carbohydrate and Glycoconjugate Analysis: Studies of Protein N-Glycosylation and Oligosaccharide Isomer Discrimination by Mass Spectrometry and Ion Mobility

Katherine N Schumacher, University of Nebraska - Lincoln

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.
http://digitalcommons.unl.edu/dissertations/AAI10793426

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