Abstract
The total cell wall mannoprotein has been isolated from a mutant of Saccharomyces cerevisiae that fails to remove the glucose units of the dolichol-linked precursor after transfer of the oligosaccharide to asparagine units in the protein. The oligosaccharides released from this mannoprotein by endoglucosaminidase H digestion show 1H NMR signals assignable to three alpha-linked glucose units as delta 5.52, 5.27, and 5.17, and a comparison with the chemical shifts of reference compounds shows that these signals are consistent with the structure alpha Glc----2 alpha Glc----3 alpha Man----2. This provides a direct confirmation for the structure previously assigned to the lipid-linked precursor. Analysis of the larger oligosaccharides confirms that the presence of the glucose units does not prevent elongation of the alpha 1----6-linked polymannose backbone or addition of alpha 1----3-linked mannose to the core.
Full text
PDFImages in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Ballou C. Structure and biosynthesis of the mannan component of the yeast cell envelope. Adv Microb Physiol. 1976;14(11):93–158. doi: 10.1016/s0065-2911(08)60227-1. [DOI] [PubMed] [Google Scholar]
- Ballou L., Cohen R. E., Ballou C. E. Saccharomyces cerevisiae mutants that make mannoproteins with a truncated carbohydrate outer chain. J Biol Chem. 1980 Jun 25;255(12):5986–5991. [PubMed] [Google Scholar]
- Cohen R. E., Ballou C. E. Linkage and sequence analysis of mannose-rich glycoprotein core oligosaccharides by proton nuclear magnetic resonance spectroscopy. Biochemistry. 1980 Sep 2;19(18):4345–4358. doi: 10.1021/bi00559a031. [DOI] [PubMed] [Google Scholar]
- Cohen R. E., Zhang W., Ballou C. E. Effects of mannoprotein mutations on Saccharomyces cerevisiae core oligosaccharide structure. J Biol Chem. 1982 May 25;257(10):5730–5737. [PubMed] [Google Scholar]
- Esmon B., Esmon P. C., Schekman R. Early steps in processing of yeast glycoproteins. J Biol Chem. 1984 Aug 25;259(16):10322–10327. [PubMed] [Google Scholar]
- Huffaker T. C., Robbins P. W. Temperature-sensitive yeast mutants deficient in asparagine-linked glycosylation. J Biol Chem. 1982 Mar 25;257(6):3203–3210. [PubMed] [Google Scholar]
- Li E., Tabas I., Kornfeld S. The synthesis of complex-type oligosaccharides. I. Structure of the lipid-linked oligosaccharide precursor of the complex-type oligosaccharides of the vesicular stomatitis virus G protein. J Biol Chem. 1978 Nov 10;253(21):7762–7770. [PubMed] [Google Scholar]
- Liu T., Stetson B., Turco S. J., Hubbard S. C., Robbins P. W. Arrangement of glucose residues in the lipid-linked oligosaccharide precursor of asparaginyl oligosaccharides. J Biol Chem. 1979 Jun 10;254(11):4554–4559. [PubMed] [Google Scholar]
- Novick P., Field C., Schekman R. Identification of 23 complementation groups required for post-translational events in the yeast secretory pathway. Cell. 1980 Aug;21(1):205–215. doi: 10.1016/0092-8674(80)90128-2. [DOI] [PubMed] [Google Scholar]
- Ogawa T., Nukada T., Kitajima T. Synthesis of a linear, hexahexosyl unit of a high-mannose type of glycan chain of a glycoprotein. Carbohydr Res. 1983 Nov 11;123(1):C12–C15. doi: 10.1016/0008-6215(83)88399-2. [DOI] [PubMed] [Google Scholar]
- Parodi A. J., Quesada-Allue L. A. Protein glycosylation in Trypanosoma cruzi. I. Characterization of dolichol-bound monosaccharides and oligosaccharides synthesized "in vivo". J Biol Chem. 1982 Jul 10;257(13):7637–7640. [PubMed] [Google Scholar]
- Raschke W. C., Kern K. A., Antalis C., Ballou C. E. Genetic control of yeast mannan structure. Isolation and characterization of mannan mutants. J Biol Chem. 1973 Jul 10;248(13):4660–4666. [PubMed] [Google Scholar]
- Runge K. W., Huffaker T. C., Robbins P. W. Two yeast mutations in glucosylation steps of the asparagine glycosylation pathway. J Biol Chem. 1984 Jan 10;259(1):412–417. [PubMed] [Google Scholar]
- Saunier B., Kilker R. D., Jr, Tkacz J. S., Quaroni A., Herscovics A. Inhibition of N-linked complex oligosaccharide formation by 1-deoxynojirimycin, an inhibitor of processing glucosidases. J Biol Chem. 1982 Dec 10;257(23):14155–14161. [PubMed] [Google Scholar]
- Tsai P. K., Frevert J., Ballou C. E. Carbohydrate structure of Saccharomyces cerevisiae mnn9 mannoprotein. J Biol Chem. 1984 Mar 25;259(6):3805–3811. [PubMed] [Google Scholar]
- Ugalde R. A., Staneloni R. J., Leloir L. F. Microsomal glucosidases of rat liver. Partial purification and inhibition by disaccharides. Eur J Biochem. 1980 Dec;113(1):97–103. doi: 10.1111/j.1432-1033.1980.tb06144.x. [DOI] [PubMed] [Google Scholar]