Enzymatic Sequence Analysis of N-Glycans by Exoglycosidase Cleavage and Mass Spectrometry: Detection of Lewis X Structures

Autor:	Christoph Kannicht, Lothar Lucka, Detlef Grunow
Rok vydání:	2019
Předmět:	chemistry.chemical_classification Glycan Chromatography chemistry biology Sequence analysis Exoglycosidase biology.protein Sequence (biology) Carbohydrate conformation Oligosaccharide Mass spectrometry Glycoprotein
Zdroj:	Post-Translational Modification of Proteins ISBN: 9781493990535 Methods in Molecular Biology™ ISBN: 9781588297198
DOI:	10.1007/978-1-4939-9055-9_5
Popis:	Enzymatic sequencing of oligosaccharides gives structural information on sequence of monosaccharides and type of linkage within the oligosaccharide chain. This data can be obtained by stepwise enzymatic digestion of a single, isolated oligosaccharide using individual or mixtures of specific exoglycosidases. N-glycans have to be fractionated from mixtures prior to sequence analysis to assign this type of structural information to a specific glycan. Enzymatic sequencing can be applied to oligosaccharide mixtures as well to evaluate the occurrence of distinct oligosac-charide motives of functional and/or structural interest. Here we describe the application of enzymatic sequence analysis to a mixture of N-glycans released from alpha1-acid glycoprotein. The experimental conditions are optimized for detection of possible Lewis X structures after stepwise exoglycosi-dase digestion by MALDI-TOF mass spectrometry. However, the described method is generally applicable to analyze other structural properties of N-glycans by use of the respective specific exoglycosidases.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_dedup___::c01d49ead24e018a2a7d8d930dba3614 https://doi.org/10.1007/978-1-4939-9055-9_5 Zobrazit plný text záznamu