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Titolo:
Nonglucosylated oligosaccharides are transferred to protein in MI8-5 Chinese hamster ovary cells
Autore:
Quellhorst, GJ; ORear, JL; Cacan, R; Verbert, A; Krag, SS;
Indirizzi:
JohnsUSApkins Univ, Sch Hyg & Publ Hlth, Dept Biochem, Baltimore, MD 21205Johns Hopkins Univ Baltimore MD USA 21205 pt Biochem, Baltimore, MD 21205 Univ Sci & Tech Lille Flandres Artois, UMR 111 CNRS, Chim Biol Lab, F-59655 Univ Sci & Tech Lille Flandres Artois Villeneuve Dascq France F-59655 55
Titolo Testata:
GLYCOBIOLOGY
fascicolo: 1, volume: 9, anno: 1999,
pagine: 65 - 72
SICI:
0959-6658(199901)9:1<65:NOATTP>2.0.ZU;2-K
Fonte:
ISI
Lingua:
ENG
Soggetto:
SACCHAROMYCES-CEREVISIAE; GLYCOPROTEIN GLUCOSYLTRANSFERASE; LINKED OLIGOSACCHARIDES; TRYPANOSOMATID PROTOZOA; GLYCOSYLATION PATHWAY; GLUCOSYLATION STEPS; N-GLYCOSYLATION; GLUCOSE; PURIFICATION; DEFICIENT;
Keywords:
ALG6; CHO cells; glucosyl transferase; N-linked glycosylation;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
38
Recensione:
Indirizzi per estratti:
Indirizzo: Krag, SS Johnsimore,ns Univ, Sch Hyg & Publ Hlth, Dept Biochem, 615 N Wolfe St, Balt Johns Hopkins Univ 615 N Wolfe St Baltimore MD USA 21205 St, Balt
Citazione:
G.J. Quellhorst et al., "Nonglucosylated oligosaccharides are transferred to protein in MI8-5 Chinese hamster ovary cells", GLYCOBIOLOG, 9(1), 1999, pp. 65-72

Abstract

A CHO mutant MI8-5 was found to synthesize Man(9)-GlcNAc(2)-P-P-dolichol rather than Glc(3)Man(9)GlcNAc(2)-P-P-dolichol as the oligosaccharide-lipid intermediate in N-glycosylation of proteins. MI8-5 cells were incubated with labeled mevalonate, and the prenol was found to be dolichol. The mannose-labeled oligosaccharide released from oligosaccharide-lipid of MI8-5 cells was analyzed by HPLC and a-mannosidase treatment, and the data were consistent with a structure of Man(9)GlcNAc(2). In addition, MI8-5 cells did not incorporate radioactivity into oligosaccharide-lipid during an incubation with tritiated galactose, again consistent with MI8-5 cells synthesizing an unglucosylated oligosaccharide-lipid, MI8-5 cells had parental levels of glucosylphosphoryldolichol synthase activity. However, in two different assays, MI8-5 cells lacked dolichol-P-Glc:Man(9)GlcNAc(2)-P-P-dolichol glucosyltransferase activity. MI8-5 cells were found to synthesize glucosylated oligosaccharide after they were transfected with Saccharomyces cerevisiae ALG6, the gene for dolichol-P-Glc:Man(9)GlcNAc(2)-P-P-dolichol glucosyltransferase. MI8-5 cells mere found to incorporate mannose into protein 2-fold slowerthan parental cells and to approximately a 2-fold lesser extent.

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Documento generato il 24/09/20 alle ore 23:18:30