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

• 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.

Ämnesord

NATURVETENSKAP  -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)

NATURAL SCIENCES  -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)

Nyckelord

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

Publikations- och innehållstyp

ref (ämneskategori)

art (ämneskategori)