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. 1985 Mar 1;226(2):487–498. doi: 10.1042/bj2260487

Hybrid sialylated N-glycans are minor constituents of normal BHK-cell glycoproteins and a prominent feature in glycoproteins of some ricin-resistant cell lines.

R C Hughes, G Mills
PMCID: PMC1144736  PMID: 3994670

Abstract

Baby-hamster kidney (BHK) cells were labelled metabolically by growth in media containing radioactive sugars and the asparagine-linked glycopeptides (N-glycans) obtained by Pronase digestion of disrupted cells were fractionated by chromatography on concanavalin A-Sepharose. About 2-3% of the total [3H]galactose- or [3H]fucose-labelled glycopeptides were found to be bound tightly to the lectin column and were eluted with 500 mM-methyl alpha-mannoside. Further analysis of these minor components by chromatography on Bio-Gel P4, lentil-lectin-Sepharose and DEAE-Sephacel and sensitivity to alpha-mannosidase indicates the presence in BHK-cell glycopeptides of hybrid structures of the following form: (Formula: see text) Similar structures were identified as major features of the glycoproteins of ricin-resistant mutants RicR17 and RicR19 as described previously for RicR21 cells [Hughes, Mills & Stojanovic (1983) Carbohydr. Res. 120, 215-234]. The RicR15 cell line also produces significant amounts of hybrid N-glycans. The studies show that the novel N-glycans accumulating in ricin-resistant mutants are derived by a metabolic pathway that exists to a minor extent in normal BHK cells.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Baenziger J. U., Fiete D. Structural determinants of Ricinus communis agglutinin and toxin specificity for oligosaccharides. J Biol Chem. 1979 Oct 10;254(19):9795–9799. [PubMed] [Google Scholar]
  2. Baenziger J. U., Fiete D. Structural determinants of concanavalin A specificity for oligosaccharides. J Biol Chem. 1979 Apr 10;254(7):2400–2407. [PubMed] [Google Scholar]
  3. Cummings R. D., Kornfeld S. Characterization of the structural determinants required for the high affinity interaction of asparagine-linked oligosaccharides with immobilized Phaseolus vulgaris leukoagglutinating and erythroagglutinating lectins. J Biol Chem. 1982 Oct 10;257(19):11230–11234. [PubMed] [Google Scholar]
  4. Cummings R. D., Kornfeld S. Fractionation of asparagine-linked oligosaccharides by serial lectin-Agarose affinity chromatography. A rapid, sensitive, and specific technique. J Biol Chem. 1982 Oct 10;257(19):11235–11240. [PubMed] [Google Scholar]
  5. Gleeson P. A., Schachter H. Control of glycoprotein synthesis. J Biol Chem. 1983 May 25;258(10):6162–6173. [PubMed] [Google Scholar]
  6. Hughes R. C., Mills G. Analysis by lectin affinity chromatography of N-linked glycans of BHK cells and ricin-resistant mutants. Biochem J. 1983 Jun 1;211(3):575–587. doi: 10.1042/bj2110575. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hughes R. C., Mills G., Stojanovic D. Hybrid, sialylated N-glycans accumulate in a ricin-resistant mutant of baby hamster kidney BHK cells. Carbohydr Res. 1983 Aug 16;120:215–234. doi: 10.1016/0008-6215(83)88018-5. [DOI] [PubMed] [Google Scholar]
  8. Hunt L. A., Wright S. E. Rous sarcoma virus glycoproteins contain hybrid-type oligosaccharides. J Virol. 1981 Aug;39(2):646–650. doi: 10.1128/jvi.39.2.646-650.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kornfeld K., Reitman M. L., Kornfeld R. The carbohydrate-binding specificity of pea and lentil lectins. Fucose is an important determinant. J Biol Chem. 1981 Jul 10;256(13):6633–6640. [PubMed] [Google Scholar]
  10. Liang C. J., Yamashita K., Muellenberg C. G., Shichi H., Kobata A. Structure of the carbohydrate moieties of bovine rhodopsin. J Biol Chem. 1979 Jul 25;254(14):6414–6418. [PubMed] [Google Scholar]
  11. Meager A., Ungkitchanukit A., Hughes R. C. Variants of hamster fibroblasts resistant to Ricinus communis toxin (ricin). Biochem J. 1976 Jan 15;154(1):113–124. doi: 10.1042/bj1540113. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Meager A., Ungkitchanukit A., Nairn R., Hughes R. C. Ricin resistance in baby hamster kidney cells. Nature. 1975 Sep 11;257(5522):137–139. doi: 10.1038/257137a0. [DOI] [PubMed] [Google Scholar]
  13. Narasimhan S., Wilson J. R., Martin E., Schachter H. A structural basis for four distinct elution profiles on concanavalin A--Sepharose affinity chromatography of glycopeptides. Can J Biochem. 1979 Jan;57(1):83–96. doi: 10.1139/o79-011. [DOI] [PubMed] [Google Scholar]
  14. Pâquet M. R., Narasimhan S., Schachter H., Moscarello M. A. Branch specificity of purified rat liver Golgi UDP-galactose: N-acetylglucosamine beta-1,4-galactosyltransferase. Preferential transfer of of galactose on the GlcNAc beta 1,2-Man alpha 1,3-branch of a complex biantennary Asn-linked oligosaccharide. J Biol Chem. 1984 Apr 25;259(8):4716–4721. [PubMed] [Google Scholar]
  15. Rosen S. W., Hughes R. C. Effects of neuraminidase on lectin binding by wild-type and ricin-resistant strains of hamster fibroblasts. Biochemistry. 1977 Nov 1;16(22):4908–4915. doi: 10.1021/bi00641a026. [DOI] [PubMed] [Google Scholar]
  16. Schachter H., Narasimhan S., Gleeson P., Vella G. Control of branching during the biosynthesis of asparagine-linked oligosaccharides. Can J Biochem Cell Biol. 1983 Sep;61(9):1049–1066. doi: 10.1139/o83-134. [DOI] [PubMed] [Google Scholar]
  17. Tulsiani D. R., Touster O. Swainsonine causes the production of hybrid glycoproteins by human skin fibroblasts and rat liver Golgi preparations. J Biol Chem. 1983 Jun 25;258(12):7578–7585. [PubMed] [Google Scholar]
  18. Vischer P., Hughes R. C. Glycosyl transferases of baby-hamster-kidney (BHK) cells and ricin-resistant mutants. N-glycan biosynthesis. Eur J Biochem. 1981 Jul;117(2):275–284. doi: 10.1111/j.1432-1033.1981.tb06334.x. [DOI] [PubMed] [Google Scholar]
  19. Yamashita K., Tachibana Y., Nakayama T., Kitamura M., Endo Y., Kobata A. Structural studies of the sugar chains of human parotid alpha-amylase. J Biol Chem. 1980 Jun 25;255(12):5635–5642. [PubMed] [Google Scholar]
  20. Yoshima H., Takasaki S., Kobata A. The asparagine-linked sugar chains of the glycoproteins in calf thymocyte plasma membrane. Structural studies of acidic oligosaccharides. J Biol Chem. 1980 Nov 25;255(22):10793–10804. [PubMed] [Google Scholar]

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