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

✍ Scribed by Mitsuhiro Kinoshita; Kohei Inagake; Atsufumi Kawabata; Ryotaro Kuroda; Yasuo Oda; Kazuaki Kakehi

• Book ID: 102563944
• Publisher: Elsevier Science
• Year: 2000
• Tongue: English
• Weight: 64 KB
• Volume: 284
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1006/abio.2000.4689

✦ Synopsis

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 E 1 analogue in vivo. Thus, the galactose oxidaseperoxidase method is useful for the determination of mucin-type glycoproteins in biological materials.