| Sialylation of N-glycans on the recombinant proteins expressed by a baculovirus-insect cell system under beta-N-acetylglucosaminidase inhibition. | |
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MedLine Citation:
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PMID: 11741890 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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We investigated the ability of a baculovirus-insect cell system to produce sialylated glycoproteins. Despite the presence of enzymes for synthesizing complex-type N-glycans, the most frequent structure of insect N-glycan is the paucimannosidic type, Man(3)GlcNAc(2)(+/-Fuc). The reason for the overwhelming assembly of paucimannosidic N-glycans is not yet well understood. We hypothesized that this predominance might be due to insect-specific, Golgi-associated beta-N-acetylglucosaminidase (GlcNAcase)-mediated removal of N-acetylglucosamine residues from the precursor N-glycan, thereby preventing its galactosylation and terminal sialylation. As we expected, the suppression of intrinsic GlcNAcase activity with a specific inhibitor, 2-acetamido-1,2-dideoxynojirimycin, allowed the accumulation of sialylated glycoproteins in the supernatants of insect cell cultures after baculoviral infection. Our observation indicates that GlcNAcase-dependent depletion of N-acetylglucosamine residues from intermediate N-glycans is critical for the assembly of paucimannosidic N-glycans in insect cells and, more importantly, that insect cells (under specific conditions) retain the ability to construct sialylated N-glycans like those in mammalian cells. |
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Authors:
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Satoko Watanabe; Takehiro Kokuho; Hitomi Takahashi; Masashi Takahashi; Takayuki Kubota; Shigeki Inumaru |
Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2001-12-06 |
Journal Detail:
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Title: The Journal of biological chemistry Volume: 277 ISSN: 0021-9258 ISO Abbreviation: J. Biol. Chem. Publication Date: 2002 Feb |
Date Detail:
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Created Date: 2002-02-11 Completed Date: 2002-03-21 Revised Date: 2007-11-15 |
Medline Journal Info:
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Nlm Unique ID: 2985121R Medline TA: J Biol Chem Country: United States |
Other Details:
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Languages: eng Pagination: 5090-3 Citation Subset: IM |
Affiliation:
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Department of Immunology, National Institute of Animal Health, 3-1-5 Kannondai, Tsukuba, Ibaraki 305-0856, Japan. satochan@affrc.go.jp |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Acetylglucosaminidase
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antagonists & inhibitors* Animals Baculoviridae Binding Sites Blotting, Western Cattle Cell Line DNA, Complementary / metabolism Enzyme Inhibitors / pharmacology Glycoproteins / chemistry*, metabolism Golgi Apparatus / enzymology Insects Lectins / metabolism Mice Models, Chemical N-Acetylneuraminic Acid / chemistry*, metabolism* Polysaccharides / metabolism* Protein Binding RNA, Messenger / metabolism Recombinant Proteins / metabolism* |
| Chemical | |
Reg. No./Substance:
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0/DNA, Complementary; 0/Enzyme Inhibitors; 0/Glycoproteins; 0/Lectins; 0/Polysaccharides; 0/RNA, Messenger; 0/Recombinant Proteins; 131-48-6/N-Acetylneuraminic Acid; EC 3.2.1.52/Acetylglucosaminidase |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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