Purification, Characterization, and Cloning of a Spodoptera Frugiperda Sf9 Β-n-acetylhexosaminidase That Hydrolyzes Terminal N-acetylglucosamine on N-glycan Core
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چکیده
PURIFICATION, CHARACTERIZATION, AND CLONING OF A SPODOPTERA FRUGIPERDA SF9 β-N-ACETYLHEXOSAMINIDASE THAT HYDROLYZES TERMINAL N-ACETYLGLUCOSAMINE ON N-GLYCAN CORE Noboru Tomiya, Someet Narang, Jung Park, Badarulhisam Abdul-Rahman, One Choi, Sundeep Singh, Jun Hiratake, Kanzo Sakata, Michael J. Betenbaugh, Karen B. Palter, Yuan C. Lee Department of Biology, Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, Maryland 21218, USA; Department of Biology, Temple University, 1900 North 12th Street, Philadelphia, PA 19122, USA; Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Yusong-gu, Taejon 305-701, Korea; Institute for Chemical Research, Kyoto University, Uji, Kyoto 611-0011, Japan Running Title: Hexosaminidase from S. frugiperda Address correspondence to: Noboru Tomiya, Department of Biology, Johns Hopkins University, 3400 North Charles Street, Baltimore, Maryland 21218, Tel. 410-516-7322; Fax. 410-516-8716: E-Mail: [email protected]
منابع مشابه
Purification, characterization, and cloning of a Spodoptera frugiperda Sf9 beta-N-acetylhexosaminidase that hydrolyzes terminal N-acetylglucosamine on the N-glycan core.
Paucimannosidic glycans are often predominant in N-glycans produced by insect cells. However, a beta-N-acetylhexosaminidase responsible for the generation of paucimannosidic glycans in lepidopteran insect cells has not been identified. We report the purification of a beta-N-acetylhexosaminidase from the culture medium of Spodoptera frugiperda Sf9 cells (Sfhex). The purified Sfhex protein showed...
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The hemagglutinin of fowl plague virus has been expressed in Spodoptera frugiperda (Sf9) cells and in Estigmene acrea cells by using a baculovirus vector. Structural analysis revealed that the endo-H-resistant N-glycans of HA from Sf9 cells were predominantly trimannosyl core oligosaccharides, whereas in E. acrea cells most of these cores were elongated by at least one terminal N-acetylglucosam...
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UDP-GlcNAc:Galbeta1-3GalNAc-R (GlcNAc to GalNAc) beta-1, 6-N-acetylglucosaminyltransferase (C2GnT) catalyses the formation of O-glycan core 2. Purification and characterization of C2GnT from natural sources has been hampered by the instability of this enzyme. We have been able to prepare a stable partly purified recombinant human C2GnT by expression of a truncated form of the enzyme in the bacu...
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Man(α1-6)[GlcNAc(β1-2)Man(α1-3)]ManGlcNAc(2) is a key branch point intermediate in the insect N-glycosylation pathway because it can be either trimmed by a processing β-N-acetylglucosaminidase (FDL) to produce paucimannosidic N-glycans or elongated by N-acetylglucosaminyltransferase II (GNT-II) to produce complex N-glycans. N-acetylglucosaminyltransferase I (GNT-I) contributes to branch point i...
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