In vitro labeling of the sialic acid moiety of glycoconjugates with carbon-14.
Labeling of sialoglycoproteins with carbon-14 in vitro was performed by reacting the aldehyde groups, generated by mild periodate oxidation of the terminal sialyl groups, with 14C-labeled sodium cyanide to produce the labeled cyanohydrin derivatives (Kiliani reaction). Labeling with tritium was carried out by reduction of the aldehyde groups generated on the sialyl residues with 3H-labeled sodium borohydride following standard procedures. The behavior of both types of labeled specimens of fetuin and ovine submaxillary mucin, individually and in mixtures, was investigated by gel-filtration chromatography, gel electrophoresis, and cesium bromide gradient ultracentrifugation. The labeled sialyl residues were subjected to partial characterization: color yield with the resorcinol and thiobarbituric acid reagents, behavior on ion-exchange chromatography, and susceptibility to mild acid and enzymatic hydrolyses. In addition to these model glycoproteins, this procedure was also utilized to label the sialoglycoproteins present in human tracheobronchial secretions collected from normal subjects and patients with chronic bronchitis. The potential uses of this approach for comparative studies of normal and pathological sialoglycoconjugates available in minute amounts is described. The extension of this approach to the labeling of the galactosyl and N-acetylgalactosaminyl moieties of glycoconjugates following treatment with galactose oxidase is outlined.
D000818:Animals; D002250:Carbon Radioisotopes; D002499:Centrifugation, Density Gradient; D002845:Chromatography / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000032:analysis*; D006868:Hydrolysis; D007553:Isotope Labeling / Q000379:methods*; D009077:Mucins / Q000032:analysis; D019158:N-Acetylneuraminic Acid; D012756:Sheep; D012794:Sialic Acids / Q000032:analysis*; D013363:Submandibular Gland / Q000032:analysis; D000509:alpha-Fetoproteins / Q000032:analysis
D002250:Carbon Radioisotopes; D006023:Glycoproteins; D009077:Mucins; D012794:Sialic Acids; D000509:alpha-Fetoproteins; D019158:N-Acetylneuraminic Acid