Structural elucidation of a variety of GalNAc-containing N-linked oligosaccharides from human urinary kallidinogenase

Fifteen different structures of terminal GalNAc-containing N-linked oligosaccharides from human urinary kallidinogenase have been identified. These N-linked oligosaccharides were mostly neutral, because sialic acid content was lower than 0.13 mol of sialic acid/mol of sugar chain, and sulfate was no...

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Published in:The Journal of biological chemistry 1993-01, Vol.268 (1), p.113-126
Main Authors: TOMIYA, N, AWAYA, J, KURONO, M, HANZAWA, H, SHIMADA, I, ARATA, Y, YOSHIDA, T, TAKAHASHI, N
Format: Article
Language:English
Subjects:
Acetylgalactosamine - analysis
Analytical, structural and metabolic biochemistry
Biological and medical sciences
Carbohydrate Conformation
Carbohydrate Sequence
Chromatography
Fundamental and applied biological sciences. Psychology
Gas Chromatography-Mass Spectrometry
Glycoproteins
Humans
Indicators and Reagents
kallidinogenase
Kallikreins - chemistry
Kallikreins - urine
Magnetic Resonance Spectroscopy
man
Molecular Sequence Data
oligosaccharides
Oligosaccharides - chemistry
Oligosaccharides - isolation & purification
Proteins
structure
urine
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Summary:Fifteen different structures of terminal GalNAc-containing N-linked oligosaccharides from human urinary kallidinogenase have been identified. These N-linked oligosaccharides were mostly neutral, because sialic acid content was lower than 0.13 mol of sialic acid/mol of sugar chain, and sulfate was not detected. The oligosaccharides were released from pepsin-digested protein by glycoamidase A (from almond) digestion. The reducing ends of the oligosaccharide chains were aminated with a fluorescent reagent, 2-aminopyridine. The resulting mixture of pyridylamino derivatives of the oligosaccharides were separated by high performance liquid chromatography on an ODS-silica column, and 15 oligosaccharides were isolated. The structure of each oligosaccharide fraction was analyzed by two-dimensional sugar mapping, component sugar analysis, high resolution proton nuclear magnetic resonance and methylation analysis. It was found that each N-linked oligosaccharide associated with human urinary kallidinogenase contains unsubstituted GalNAc residues at the nonreducing terminal. These 15 oligosaccharides include 5 biantennary, 7 triantennary, and 3 tetraantennary oligosaccharides.
ISSN:0021-9258
1083-351X
DOI:10.1016/S0021-9258(18)54122-3