Document Detail


Sialylation of N-glycans on the recombinant proteins expressed by a baculovirus-insect cell system under beta-N-acetylglucosaminidase inhibition.
MedLine Citation:
PMID:  11741890     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Satoko Watanabe; Takehiro Kokuho; Hitomi Takahashi; Masashi Takahashi; Takayuki Kubota; Shigeki Inumaru
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2001-12-06
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  277     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  2002 Feb 
Date Detail:
Created Date:  2002-02-11     Completed Date:  2002-03-21     Revised Date:  2007-11-15    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  5090-3     Citation Subset:  IM    
Affiliation:
Department of Immunology, National Institute of Animal Health, 3-1-5 Kannondai, Tsukuba, Ibaraki 305-0856, Japan. satochan@affrc.go.jp
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MeSH Terms
Descriptor/Qualifier:
Acetylglucosaminidase / 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:
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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