A concanavalin A-resistant Chinese hamster ovary cell line is deficient in the synthesis of [3H]glucosyl oligosaccharide-lipid

In this report we present an initial determination of the biochemical defect present in a Chinese hamster ovary cell line selected for resistance to concanavalin A. Membranes of this mutant, B211, incorporated at least 10-fold less mannose from GDP-[14C]mannose into oligosaccharide-lipid than membra...

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Bibliographic Details
Published in:The Journal of biological chemistry 1979-09, Vol.254 (18), p.9167-9177
Main Author: Krag, S S
Format: Article
Language:English
Subjects:
Animals
Cell Line
Cell Membrane - metabolism
Concanavalin A - pharmacology
Cricetinae
Female
Glucose - metabolism
Glycolipids - biosynthesis
Guanosine Diphosphate Mannose - metabolism
Kinetics
Mutation
Oligosaccharides - biosynthesis
Ovary
Polyisoprenyl Phosphate Sugars - metabolism
Uridine Diphosphate N-Acetylglucosamine - metabolism
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Summary:In this report we present an initial determination of the biochemical defect present in a Chinese hamster ovary cell line selected for resistance to concanavalin A. Membranes of this mutant, B211, incorporated at least 10-fold less mannose from GDP-[14C]mannose into oligosaccharide-lipid than membranes of the wild type. In the presence of dolichol phosphate, membranes of the mutant and wild type exhibited similar rates of synthesis of number of early intermediates, namely, mannosylphosphoryldolichol, N-acetylglucosaminyl- and N,N'-diacetylchitobiosylpyrophosphoryldolichol, glucosylphosphoryldolichol, and mannosyloligosaccharide-lipid. The membranes of B211 did not incorporate glucose from UDP-[3H]glucose into oligosaccharide-lipid or protein. Comparison by gel filtration chromatography of oligosaccharides derived from the oligosaccharide-lipids of B211 and wild type cells labeled with [2-3H]mannose revealed that B211 cells incorporated little if any label into an oligosaccharide corresponding to the most excluded oligosaccharide labeled by wild type cells. This concanavalin A-resistant cell line appears to lack the ability to glucosylate oligosaccharide-lipid.
ISSN:0021-9258
1083-351X
DOI:10.1016/S0021-9258(19)86826-6