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 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.
Natsuka
S
S
Department of Chemistry, Osaka University College of Science.
Himeno
M
M
Hase
S
S
Ito
H
H
Ueda
T
T
Kato
K
K
Ikenaka
T
T
D000818:Animals; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002241:Carbohydrates / Q000032:analysis*; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004950:Esterases / Q000032:analysis; D006026:Glycoside Hydrolases; D006834:Hydrazines; D009682:Magnetic Resonance Spectroscopy; D008862:Microsomes, Liver / Q000201:enzymology*; D008969:Molecular Sequence Data; D009596:Nitrophenols / Q000378:metabolism*; D011487:Protein Conformation; D051381:Rats
D002241:Carbohydrates; D006834:Hydrazines; D009596:Nitrophenols; C029424:hydrazine; C008642:4-nitrophenyl acetate; D004950:Esterases; D006026:Glycoside Hydrolases