The Carbohydrate Moieties of Human Urinary Ribonuclease UL

Ribonuclease UL. purified from pooled human urine contains approximately 20.7% of neutral sugar and 7.8% of aminosugar. All sugars were quantitatively released as oligosaccharides on hydrazinolysis. The oligosaccharides were converted to tritium-labeled oligosaccharides on reduction with NaB3H4 The...

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Bibliographic Details
Published in:Journal of biochemistry (Tokyo) 1987-01, Vol.101 (1), p.29-41
Main Authors: HITOI, Akira, YAMASHITA, Katsuko, NIWATA, Yasushi, IRIE, Masachika, KOCHIBE, Naohisa, KOBATA, Akira
Format: Article
Language:English
Subjects:
Analytical, structural and metabolic biochemistry
Biological and medical sciences
Carbohydrates - analysis
Chromatography, Ion Exchange
Chromatography, Paper
Enzymes and enzyme inhibitors
Fundamental and applied biological sciences. Psychology
Galactose - analysis
Glycoside Hydrolases - metabolism
Humans
Hydrolases
Hydrolysis
Oligosaccharides - analysis
Ribonucleases - urine
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Summary:Ribonuclease UL. purified from pooled human urine contains approximately 20.7% of neutral sugar and 7.8% of aminosugar. All sugars were quantitatively released as oligosaccharides on hydrazinolysis. The oligosaccharides were converted to tritium-labeled oligosaccharides on reduction with NaB3H4 The radioactive oligosaccharide fraction was separated into a neutral and an acidic fraction on paper electrophoresis. All oligosaccharides in the acidic fraction could be converted to neutral oligosaccharides with the release of one sialic acid residue by sialidase digestion. Both fractions were shown to be mixtures of more than fourteen oligosaccharides by gel permeation chromatography. Structural studies on these oligosaccharides involving sequential exoglycosidase digestion in combination with methylation analysis revealed that ribonuclease UL. contains sialylated and nonsialylated mono, bi-, tri-, and tetraantennary complex type sugar chains with N acetyllactosamine outer chains, and tri- and tetraantennary complex type sugar chains with various numbers of Galβ1→4GlcNAcβ1 → 3Galfll→4GlcNAcfll→Outer chains. An important finding was that all sialic acid residues in the acidic oligosaccharides only occur as the Siaa2→6Galβ1 →4GlcNAcβ1→2Mana1→3 group. Both fucosylated and non-fucosylated trimannosyl cores were found among the asparagine-linked sugar chains of ribonuclease UL.
ISSN:0021-924X
1756-2651
DOI:10.1093/oxfordjournals.jbchem.a121905