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 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.
Tomiya
N
N
Mie Research Laboratory, Sanwa Kagaku Kenkyusho Co. Ltd., Japan.
Awaya
J
J
Kurono
M
M
Hanzawa
H
H
Shimada
I
I
Arata
Y
Y
Yoshida
T
T
Takahashi
N
N
D000116:Acetylgalactosamine / Q000032:analysis*; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002845:Chromatography; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D007202:Indicators and Reagents; D007610:Kallikreins / Q000737:chemistry / Q000652:urine*; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry* / Q000302:isolation & purification
D007202:Indicators and Reagents; D009844:Oligosaccharides; D007610:Kallikreins; D000116:Acetylgalactosamine