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The primary glycosylation defect in class e thy 1 negative mutant mouse lymphoma cells is an inability to synthesize dolichol phospho mannose

, : The primary glycosylation defect in class e thy 1 negative mutant mouse lymphoma cells is an inability to synthesize dolichol phospho mannose. Journal of Biological Chemistry 255(10): 4441-4446

Thy-1-- mutant mouse lymphoma cells of the class E complementation group are unable to synthesize the normal Glc3Man9GlcNAc2 lipid-linked oligosaccharide, but instead accumulate a smaller lipid-linked species with the structure Man.alpha.1 .fwdarw. 2Man.alpha.1 .fwdarw. 2Man.alpha.1 .fwdarw. 3(Man.alpha.1 .fwdarw. 6)Man.beta.1 .fwdarw. 4GlcNAc.beta.1 .fwdarw. 4GlcNAc. The primary defect in the Thy-1- cells is an inability to synthesize dolichol-P-mannose. Intact Thy-1- cells incubated with [2-3H]mannose failed to incorporate any detectable radioactiVity into dolicho-P-mannose and crude membrane preparations of Thy-1- cells were unable to transfer mannose from GDP-[3H]mannose to dolichol-P. These membrane preparations did incorporate [3H]mannose into lipid-linked oligosaccharides up to the size of Man5GlcNAc2, suggesting that these species are formed from mannosyl donors other than dolichol-P-mannose. When class E Thy-1- membrane preparations were incubated with exogenous dolicho-P-[3H]mannose, lipid-linked oligosaccharides ranging in size from Man6GlcNAc2 to Man9GlcNAc2 were formed. The mutant cells have the .alpha.1,3-mannosyltransferase necessary for the conversion of the Man5GlcNAc2 species to Man6GlcNAc2, but lack the appropriate mannosyl donor (dolichol-P-mannose) necessary for the formation of the larger lipid-linked oligosaccharides. At least 2 mannosyl donors are involved in the synthesis of lipid-linked oligosaccharides. GDP-mannose is the probable donor for the formation of the Man1-5GlcNAc2 species, while dolichol-P-mannose donates the 6th mannosyl residue and probably mannose residues 7 through 9.

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Related references

Chapman, A.; Fujimoto, K.; Kornfeld, S., 1980: 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 255(10): 4441-4446

Kean, E.L.; DeBrakeleer, D.J., 1986: Stimulation by dolichol phosphate-mannose of N-acetylglucosaminyl-lipid biosynthesis by membranes from class E Thy-1-negative mutant mouse lymphoma cells which are defective in dolichol phosphate-mannose biosynthesis. Dolichol phosphate-mannose has been shown previously to stimulate the biosynthesis of N-acetylglucosaminyl-diphosphate-dolichol (E. L. Kean (1985) J. Biol. Chem. 260, 12561-12571). Although the class E Thy-1-negative mutant mouse lymphoma cells ar...

Kean E.L.; Debrakeleer D.J., 1986: Effect of dolichol p mannose on n acetylglucosamine lipid synthesis by membranes from class e thy 1 negative mutant mouse lymphoma cells. Federation Proceedings 45(6): 1678

Kornfeld, S.; Gregory, W.; Chapman, A., 1979: Class E Thy-1 negative mouse lymphoma cells utilize an alternate pathway of oligosaccharide processing to synthesize complex-type oligosaccharides. Journal of Biological Chemistry 254(22): 11649-11654

Larriba, G.; Elorza, M.V.; Villanueva, J.R.; Sentandreu, R., 1976: Participation of dolichol phospho-mannose in the glycosylation of yeast wall manno-proteins at the polysomal level. Febs Letters 71(2): 316-320

Yagodnik C.; D.L.C.nal L.; Parodi A.J., 1987: Tetrahymena pyriformis cells are deficient in all mannose p dolichol dependent mannosyltransferases but not in mannose p dolichol synthesis. Cells of the ciliated protozoan Tetrahymena pyriformis incubated with [14C]glucose were found to synthesize Man-P-dolichol and Glc-P-dolichol, as well as Glc3Man5GlcNAc2-P-P-dolichol, the latter being the main and largest lipid derivative formed....

Rimoldi, D.; Creek, K.E.;, L.M., 1990: Reduced mannose incorporation into GDP-mannose and dolichol-linked intermediates of N-glycosylation in hamster liver during vitamin A deficiency. The molecular mechanism of reduced incorporation of radioactively labeled mannose into hamster liver glycoconjugates during the progression of vitamin A deficiency was investigated. In particular the in vivo incorporation of [2-3H]mannose into GDP...

Poorman P.A.; Krehl R.; Turner N.T.; Clive D., 1986: Comparison of sigma tk negative negative mutant frequency and cytogenetic damage in l 5178y tk positive negative mouse lymphoma cells. Environmental Mutagenesis 8(SUPPL 6): 65-66

Teng M H.; Basch R.; Buxbaum J.N., 1987: Glycosylation defect in a murine thy 1 negative lymphoma cell line. Journal of Cellular Biochemistry Supplement (11 PART D): 156

Friedrich, U.; Coffino, P., 1977: Hypo xanthine guanine phospho ribosyl transferase ec with altered substrate affinity in mutant mouse lymphoma cells. Cells with altered hypoxanthine-guanine phosphoribosyl transferase (HPRT) (IMP:pyrophosphate phosphoribosyltransferase, EC were selected. Compared to wild type [S49 mouse lymphoma], mutant enzyme has a reduced affinity for the substrate p...