Fluorometric Determination of Mucin-Type Glycoproteins by the Galactose Oxidase-Peroxidase Method

Author： Kinoshita M. Inagake K. Kawabata A. Kuroda R. Oda Y. Kakehi K.

Publisher： Elsevier

ISSN： 0003-2697

Source： Analytical Biochemistry, Vol.284, Iss.1, 2000-08, pp. : 87-92

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Abstract

We developed a convenient and specific method for the determination of mucin-type glycoproteins using galactose oxidase and horseradish peroxidase on the basis of the contents of galactosyl and N-acetylgalactosaminyl residues in glycoproteins. Galactose and galactosamine residues released from glycoproteins after hydrolysis were oxidized with galactose oxidase and subsequently the resultant hydrogen peroxide was determined by a combination of horseradish peroxidase and 3-(p-hydroxyphenyl) propionic acid as a fluorogenic substrate. The contents of galactose/galactosamine residues in N- and O-glycans, as determined by the galactose oxidase-peroxidase method, were in good agreement with those described in the previous reports. We applied the present method to determine mucin-type glycoproteins secreted from rat gastric mucosa by stimulation with misoprostol, a prostaglandin E₁ analogue in vivo. Thus, the galactose oxidase-peroxidase method is useful for the determination of mucin-type glycoproteins in biological materials.