Mass spectrometry of glycans

Abstract

Powerful new strategies based on mass spectrometry are revolutionizing the structural analysis and profiling of glycans and glycoconjugates. We survey here the major biosynthetic pathways that underlie the biological diversity in glycobiology, with emphasis on glycoproteins, and the approaches that can be used to address the resulting heterogeneity. Included among these are derivatizations, on- and off-line chromatography, electrospray and matrix-assisted laser desorption/ionization, and a variety of dissociation methods, the recently introduced electron-based techniques being of particular interest.

Characteristic Changes in Cell Surface Glycosylation Accompany Intestinal Epithelial Cell (IEC) Differentiation: High Mannose Structures Dominate the Cell Surface Glycome of Undifferentiated Enterocytes

“…Recent advancements in mass spectrometry have overcome the limitations inherent to earlier glycan profiling methodologies (32)(33)(34)(35)(36)(37)(38)(39)(40)(41)(42)(43)(44). Herein we employ an MS-based analytical approach utilizing nano-LC separation with high resolution TOF MS for accurate detection of compounds, enabling rapid identification and quantitation of N-glycan alterations during Caco-2 cell differentiation.…”

Construction of a Database of Collision Cross Section Values for Glycopeptides, Glycans, and Peptides Determined by IM-MS

“…In our experiments, we first studied the RNase B tryptic glycopeptide 34 N*LTK 37 (where N* = occupied glycosylation site). Figure 1 displays a 2-D plot of m/z vs drift time over the drift time range during which the Man 5–9 [M+2H] 2+ ions were observed.…”

“…A two-dimensional plot displaying m/z as a function of drift time represented with a false color scale (least intense features in blue; most intense in red) for the RNase B tryptic glycopeptide 34 N*LTK 37 . To the left and above the 2-D plot are the mass and drift time spectra, respectively, for Man 5–9 [M+2H] 2+ glycopeptide ions.…”

“…31,32 Aberrant glycosylation associated with cancer, neurodegenerative disorders, and genetic abnormalities reflects disease physiology, 33–35 and many infectious agents show specificity for particular cell surface glycans. 36 Thus, essential physiological functions including cell–cell and cell–matrix recognition, 37 cellular adhesion, 38 and inter- and intracellular interactions depend on protein–carbohydrate interactions. 38 …”

“…41 Each glycoprotein is expressed as a set of glycoforms that share a common amino acid sequence as the backbone but differ in the localizations, structures, or sequences of carbohydrate units. 37 …”