A novel glycosylation phenotype expressed by Lec23, a Chinese hamster ovary mutant deficient in alpha-glucosidase I.
作者： M.K. Ray , J. Yang , S. Sundaram , P. Stanley
DOI: 10.1016/S0021-9258(18)54427-6
关键词:
摘要: Lec23 Chinese hamster ovary (CHO) cells have been shown to possess a unique lectin resistance phenotype and genotype compared with previously isolated CHO glycosylation mutants (Stanley, P., Sallustio, S., Krag, S. Dunn, B. (1990) Somatic Cell Mol. Genet. 16, 211-223). In this paper, biochemical basis for the lec23 mutation is identified. The carbohydrates associated G glycoprotein of vesicular stomatitis virus (VSV) grown in (Lec23/VSV) were found predominantly oligomannosyl that bound strongly concanavalin A-Sepharose, eluted 3 sugar eq beyond Man9GlcNAc marker oligosaccharide on ion suppression high pressure liquid chromatography, susceptible digestion jack bean alpha-mannosidase. Monosaccharide analyses revealed contained glucose, indicating defect alpha-glucosidase activity. This was confirmed by further structural characterization Lec23/VSV using purified rat mammary gland I, alpha-mannosidase, 1H NMR spectroscopy at 500 MHz. [3H]Glucose-labeled Glc3Man9GlcNAc prepared from CHO/VSV labeled [3H]galactose presence processing inhibitors castanospermine deoxymannojirimycin. Subsequently, [3H]Glc2Man9GlcNAc I [3H]Glc3Man9GlcNAc. When these oligosaccharides used as substrates it are specifically defective deficiency not identified among mammalian cell mutants.
elsevier.com
sci-hub.st 参考文章(41)
S Kornfeld, I Tabas, The synthesis of complex-type oligosaccharides. III. Identification of an alpha-D-mannosidase activity involved in a late stage of processing of complex-type oligosaccharides. Journal of Biological Chemistry. ,vol. 253, pp. 7779- 7786 ,(1978) , 10.1016/S0021-9258(17)34437-X
P A Romero, A Herscovics, Transfer of nonglucosylated oligosaccharide from lipid to protein in a mammalian cell. Journal of Biological Chemistry. ,vol. 261, pp. 15936- 15940 ,(1986) , 10.1016/S0021-9258(18)66656-6
R B Trimble, P H Atkinson, Structure of yeast external invertase Man8-14GlcNAc processing intermediates by 500-megahertz 1H NMR spectroscopy. Journal of Biological Chemistry. ,vol. 261, pp. 9815- 9824 ,(1986) , 10.1016/S0021-9258(18)67588-X
Mark R. Hardy, Monosaccharide analysis of glycoconjugates by high-performance anion-exchange chromatography with pulsed amperometric detection. Methods in Enzymology. ,vol. 179, pp. 76- 82 ,(1989) , 10.1016/0076-6879(89)79115-1
Monty Krieger, Pranhitha Reddy, Karen Kozarsky, David Kingsley, Lawrence Hobbie, Marsha Penman, Analysis of the synthesis, intracellular sorting, and function of glycoproteins using a mammalian cell mutant with reversible glycosylation defects. Methods in Cell Biology. ,vol. 32, pp. 57- 84 ,(1989) , 10.1016/S0091-679X(08)61167-X
Eve Barak Briles, Lectin-Resistant Cell Surface Variants of Eukaryotic Cells International Review of Cytology. ,vol. 75, pp. 101- 165 ,(1982) , 10.1016/S0074-7696(08)61003-7
A. Chapman, K. Fujimoto, S. Kornfeld, The primary glycosylation defect in class E Thy-1-negative mutant mouse lymphoma cells is an inability to synthesize dolichol-P-mannose. Journal of Biological Chemistry. ,vol. 255, pp. 4441- 4446 ,(1980) , 10.1016/S0021-9258(19)85510-2
M F Verostek, P H Atkinson, R B Trimble, Structure of Saccharomyces cerevisiae alg3, sec18 mutant oligosaccharides. Journal of Biological Chemistry. ,vol. 266, pp. 5547- 5551 ,(1991) , 10.1016/S0021-9258(19)67629-5
B Saunier, R D Kilker, J S Tkacz, A Quaroni, A Herscovics, Inhibition of N-linked complex oligosaccharide formation by 1-deoxynojirimycin, an inhibitor of processing glucosidases. Journal of Biological Chemistry. ,vol. 257, pp. 14155- 14161 ,(1982) , 10.1016/S0021-9258(19)45358-1
M L Reitman, I S Trowbridge, S Kornfeld, A lectin-resistant mouse lymphoma cell line is deficient in glucosidase II, a glycoprotein-processing enzyme. Journal of Biological Chemistry. ,vol. 257, pp. 10357- 10363 ,(1982) , 10.1016/S0021-9258(18)34027-4