Structures of sugar chains of a p-nitrophenyl acetate-hydrolyzing esterase from the microsomes of rat liver

Shunji Natsuka, Masaru Himeno, Sumihiro Hase, Harunobu Ito, Tadashi Ueda, Keitaro Kato, Tokuji Ikenaka

Research output: Contribution to journalArticle

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Abstract

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.

Original languageEnglish
Pages (from-to)986-991
Number of pages6
JournalJournal of biochemistry
Volume103
Issue number6
DOIs
Publication statusPublished - Jan 1 1988

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Acetylesterase
Liver Microsomes
Esterases
Sugars
Liver
Rats
High Pressure Liquid Chromatography
Glycoside Hydrolases
Asparagine
Glucosamine
Acetylation
Mannose
Gel Chromatography
Digestion
Magnetic Resonance Spectroscopy
Enzymes
4-nitrophenyl acetate
Nuclear magnetic resonance spectroscopy
Gels
alpha-aminopyridine

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Cite this

Structures of sugar chains of a p-nitrophenyl acetate-hydrolyzing esterase from the microsomes of rat liver. / Natsuka, Shunji; Himeno, Masaru; Hase, Sumihiro; Ito, Harunobu; Ueda, Tadashi; Kato, Keitaro; Ikenaka, Tokuji.

In: Journal of biochemistry, Vol. 103, No. 6, 01.01.1988, p. 986-991.

Research output: Contribution to journalArticle

Natsuka, Shunji ; Himeno, Masaru ; Hase, Sumihiro ; Ito, Harunobu ; Ueda, Tadashi ; Kato, Keitaro ; Ikenaka, Tokuji. / Structures of sugar chains of a p-nitrophenyl acetate-hydrolyzing esterase from the microsomes of rat liver. In: Journal of biochemistry. 1988 ; Vol. 103, No. 6. pp. 986-991.
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AB - 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.

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