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Higher Energy Collisional Dissociation Mass Spectrometry of Sulfated O-Linked Oligosaccharides

Journal article
Authors Samah Issa
Varvara Vitiazeva
Catherine A Hayes
Niclas G. Karlsson
Published in Journal of Proteome Research
Volume 17
Issue 9
Pages 3259-3267
ISSN 1535-3893
Publication year 2018
Published at Institute of Biomedicine, Department of Medical Biochemistry and Cell Biology
Pages 3259-3267
Language en
Keywords glycosylation, sulfation, Orbitrap, mucin, fragmentation, sulfoglycomics, glycomics, O-linked, sialyl-lewis-x, permethylated glycans, high-throughput, siglec-f, mucin, ligand, ms/ms, glycomics, Biochemistry & Molecular Biology
Subject categories Cell and molecular biology


Sulfation is the final decoration of mucin-type O-linked oligosaccharides before mucins are released into the lumen of the gastrointestinal, respiratory, and genital tracts. Because only a fraction of oligosaccharides undergo this type of modifications in the Golgi apparatus, sometimes also only by dedicated cells, the glycobiology of these low abundant sulfated oligosaccharides is often overlooked. At the same time, the technology to consistently identify and characterize them has been lagging. We adopted higher energy collisional dissociation to characterize sulfated oligosaccharides from porcine gastric and human salivary MUC5B mucins. With this approach we could generate conclusive spectra up to nonasaccharides. Both singly and doubly sulfated oligosaccharides were characterized. By comparing the fragmentation of low-mass fragments of m/z 100-320 with standards for six-linked and three linked sulfate, it could be shown that characteristic fragmentation exists, verifying that porcine gastric mucin contains mostly six linked sulfate to GlcNAc, whereas human MUC5B contains mostly three-linked Gal. When performing ion-trap MS2 fragmentation, these low-molecular-mass fragments are usually not detected. Hence it can be concluded that to be able to address biological questions of sulfation low-mass fragments are important for the assignment of sulfate position.

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