| Transfer of two oligosaccharides to protein in a Chinese hamster ovary cell B211 which utilizes polyprenol for its N-linked glycosylation intermediates. | |
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MedLine Citation:
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PMID: 9784244 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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B211, a glycosylation mutant isolated from Chinese hamster ovary cells, synthesizes 10- to 15-fold less Glc3Man9GlcNAc2-P-P-lipid, the substrate used by the oligosaccharide transferase in the synthesis of asparagine-linked glycoproteins. B211 cells are also 10- to 15-fold deficient in the glucosylation of oligosaccharide-lipid. Despite these properties, protein glycosylation in B211 cells proceeds at a level similar to (50% of) parental cells. We asked whether the near wild-type level of glycosylation was due to the transfer of alternative oligosaccharide structures to protein in B211 cells. The aberrant size of [35S]methionine-labeled VSV G protein and the increased percentage of endoglycosidase H-resistant tryptic peptides as compared to parental cells supported this hypothesis. B211 cells were labeled with [2-3H]mannose either for 1 min or for 1 h in the presence of glycoprotein-processing inhibitors so that the oligosaccharides initially transferred to protein could be analyzed. In addition to Glc3Man9GlcNAc2, a second, endoglycosidase H-resistant oligosaccharide was transferred whose structure was determined by alpha-mannosidase digestion, gel filtration chromatography, and HPLC to be Glc0,1Man5GlcNAc2. Finally, since the synthesis of reduced amounts of Glc3Man9GlcNAc2-P-P-lipid was also a phenotype seen in another glycosylation mutant, Lec9, we analyzed the long-chain prenol in B211 cells. B211 cells synthesized and utilized polyprenol rather than dolichol for all N-linked glycosylation intermediates as determined by HPLC analysis of [3H]mevalonate-labeled lipids. Cell fusions analyzed by similar techniques indicated that B211, originally isolated as a concanavalin A-resistant cell line, is in the Lec9 complementation group. |
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Authors:
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A Kaiden; A G Rosenwald; R Cacan; A Verbert; S S Krag |
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Publication Detail:
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Type: Journal Article; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
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Title: Archives of biochemistry and biophysics Volume: 358 ISSN: 0003-9861 ISO Abbreviation: Arch. Biochem. Biophys. Publication Date: 1998 Oct |
Date Detail:
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Created Date: 1998-11-13 Completed Date: 1998-11-13 Revised Date: 2007-11-14 |
Medline Journal Info:
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Nlm Unique ID: 0372430 Medline TA: Arch Biochem Biophys Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 303-12 Citation Subset: IM |
Copyright Information:
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Copyright 1998 Academic Press. |
Affiliation:
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School of Hygiene and Public Health, The Johns Hopkins University, Baltimore, Maryland, 21205, USA. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals CHO Cells Cricetinae Dolichol / biosynthesis Fatty Alcohols / metabolism* Glucose / metabolism Glycosylation Lipid Metabolism Lipids / biosynthesis Mannose / metabolism Molecular Sequence Data Oligosaccharides / metabolism* Proteins / metabolism* Tritium |
| Grant Support | |
ID/Acronym/Agency:
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R01 CA20421/CA/NCI NIH HHS; T32 CA09110/CA/NCI NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Fatty Alcohols; 0/Lipids; 0/Oligosaccharides; 0/Proteins; 10028-17-8/Tritium; 2067-66-5/Dolichol; 31103-86-3/Mannose; 50-99-7/Glucose |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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