Development and Application of High Performance Liquid Chromatography Map of Glucuronyl N-glycans

Abstract

Although the multi-dimensional HPLC maps of neutral, sialyl, and sulfated N-glycans have been reported and widely used for glycosylation profiling, those of glucuronyl oligosaccharides have not yet been available. In the present study, by in vitro enzymatic reactions, we prepared 55 different glucuronyl PA-oligosaccharides that include 6 kinds of HNK-1-containing N-glycans, and established their HPLC map. Furthermore, we applied this map to the characterization of branch specificity in glucuronylation reaction catalyzed by human GlcAT-S, revealing that this enzyme transfers the glucuronyl residues preferentially onto the Gal 1 4GlcNAc 1 4Man 1 3 and Gal 1 4GlcNAc 1 2Man 1 3 branches of a galactose-terminated tri-antennary oligosaccharide. The HPLC map developed in the present study will be a useful glycomics tool for identification and profiling of glucuronyl N-glycans expressed in the neural and other biological systems.Keywords: Glucuronyl oligosaccharides/N-glycans/HPLC map/GlcAT-S/branch specificity.

Development and Application of Glycosylation-Profiling Techniques for Functional Glycomics in the Nervous System

“…During evolution, the nervous system of higher animals is and TMEM5 has also been reported to be as enzymes involved in the biosynthesis of laminin binding glycans (16)(17)(18)(19). These enzymes are involved in tandem ribitol phosphate modifications linking the basal glycan moiety (GalNAc-β3-GlcNAc-β4-Man-6-phosphate core) and the outer laminin-binding region (24)(25)(26)(27).…”

Analysis of carbohydrates and glycoconjugates by matrix‐assisted laser desorption/ionization mass spectrometry: An update for 2009–2010

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Multidimensional HPLC mapping method for the structural analysis of anionic N-glycans

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