Structures of Sugar Chains of a p-Nitrophenyl Acetate-Hydrolyzing Esterase from the Microsomes of Rat Liver

The structures of sugar chains of a p-nitrophenyl acetate-hydrolyzing esterase from the microsomes of rat liver were established. The enzyme contained mannose and glucosamine as sugar components. Asparagine-linked sugar chains of the esterase were liberated by hydrazinolysis. After N-acetylation of...

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Bibliographic Details
Published in:Journal of biochemistry (Tokyo) 1988, Vol.103 (6), p.986-991
Main Authors: Natsuka, Shunji, Himeno, Masaru, Hase, Sumihiro, Ito, Harunobu, Ueda, Tadashi, Kato, Keitaro, Ikenaka, Tokuji
Format: Article
Language:English
Subjects:
Analytical, structural and metabolic biochemistry
Animals
Biological and medical sciences
Carbohydrate Conformation
Carbohydrate Sequence
Carbohydrates - analysis
Chromatography, Gel
Chromatography, High Pressure Liquid
Enzymes and enzyme inhibitors
esterase
Esterases - analysis
Fundamental and applied biological sciences. Psychology
Glycoside Hydrolases
Hydrazines
Hydrolases
liver
Magnetic Resonance Spectroscopy
Microsomes, Liver - enzymology
Molecular Sequence Data
Nitrophenols - metabolism
Protein Conformation
Rats
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Summary:The structures of sugar chains of a p-nitrophenyl acetate-hydrolyzing esterase from the microsomes of rat liver were established. The enzyme contained mannose and glucosamine as sugar components. Asparagine-linked sugar chains of the esterase were liberated by hydrazinolysis. After N-acetylation of the hydrazinolysate, the reducing ends of the sugar chains were coupled with 2-aminopyridine. Fluorescent pyridylamino (PA-) derivatives of sugar chains thus obtained were purified by gel filtration and reversed-phase HPLC. Eleven PA-sugar chains were obtained. The structures of the PA-sugar chains were first identified by HPLC using two series of separation systems by which 11 PA-oligomannose-type sugar chains with known structures could be separated. Further elucidation of the structures of each PA-sugar chain was performed by exoglycosidase digestions and partial acetolysis. The structures of two of the PA-sugar chains were further confirmed by 500 MHz 1H-NMR spectroscopy.
ISSN:0021-924X
1756-2651
DOI:10.1093/oxfordjournals.jbchem.a122398