Method for the Comparative Glycomic Analyses of O-Linked, Mucin-Type Oligosaccharides

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Abstract

A method is presented for the direct relative quantitation of distinct O-linked mucin-type oligosaccharides. Mucin-type oligosaccharides are found in a host of tissues from anuran to humans. Because they are often associated with extracellular matrix, they play important roles in cell-cell recognition. Changes in glycosylation of O-linked oligosaccharides are associated with diseases such as cancer. In fertilization, they play an active role in sperm-egg recognition. We describe a method for observing changes in glycosylation of mucin-type oligosaccharides by incorporating deuterium in the release procedure. Oligosaccharides from two different sources are released separately by sodium tetrahydroborate and sodium tetradeuterioborate. The oligosaccharides are combined and separated into components by HPLC. By observing the ratio of deuterated and undeuterated species, changes in glycosylation are precisely quantified. This method is illustrated with mucin-type oligosaccharides from the egg jelly coat of the anuran Xenopus laevis.

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