1887

Abstract

Specific labelling of the plasma membrane of intact protoplasts from , using lactoperoxidase-catalysed iodination, has permitted the development of a procedure for isolating relatively pure preparations of this component. The specific activity of the labelled membrane was very low, indicating that only a few proteins are exposed on the outer membrane surface. The specific activity of labelling increased almost 100-fold when both membrane surfaces were exposed to iodination. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis, in combination with autoradiography, indicated that only two glycosylated proteins are exposed on the outer membrane surface, whilst all membrane proteins can be labelled in isolated plasma membrane preparations. Extraction of mannan-protein from purified cell walls of gave material which, on sodium dodecyl sulphate-polyacrylamide gel electrophoresis, resembled the exposed plasma membrane glycoproteins.

Purified plasma membranes incorporated mannose from GDPmannose on to an endogenous protein acceptor. The addition of exogenous cell wall mannan-protein increased the degree of incorporation. Approximately 12 % of the incorporated mannose was sensitive to -elimination, but a lipid intermediate was not involved in the reaction. An Arrhenius plot of enzyme activity at different temperatures showed a discontinuity at 17 ° C. The mannan synthetase activity was also significantly less at 40 ° C than at 37 ° C, temperatures at which the yeast-mycelial transition has been found to occur in this organism. The possible biosynthetic relationship between the exposed glycoproteins of the plasma membrane and those of the cell wall is discussed.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-103-1-51
1977-11-01
2021-10-28
Loading full text...

Full text loading...

/deliver/fulltext/micro/103/1/mic-103-1-51.html?itemId=/content/journal/micro/10.1099/00221287-103-1-51&mimeType=html&fmt=ahah

References

  1. Ballou C. E. 1974; Some aspects of the structure, immunochemistry and genetic control of yeast mannans. Advances in Enzymology 40:239–270
    [Google Scholar]
  2. Chattaway F. W., Holmes M. R., Barlow A. J. E. 1968; Cell wall composition of the mycelial and blastospore forms of Candida albicans. Journal of General Microbiology 51:367–376
    [Google Scholar]
  3. Douglas L. J., Atkinson D. M., Hossack J. A., Rose A. H. 1975; A possible role for lipid unsaturation in the biosynthesis of wall polymers in Saccharomyces cerevisiae. Abstracts, Fourth International Symposium on Yeast and Other Protoplasts p. 71 Leicester: Cave Printing Group;
    [Google Scholar]
  4. Duran A., Bowers B., Cabib E. 1975; Chitin synthetase zymogen is attached to the yeast plasma membrane. Proceedings of the National Academy of Sciences of the United States of America 72:3952–3955
    [Google Scholar]
  5. Fairbanks G., Steck T. L., Wallach D. F. H. 1971; Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry 10:2606–2617
    [Google Scholar]
  6. Falcone G., Nickerson W. J. 1956; Cell wall mannan-protein of Baker’s yeast. Science 124:272–273
    [Google Scholar]
  7. Gold M. H., Hahn H. J. 1976; Role of a mannosyl lipid intermediate in the synthesis of Neurospora crassa glycoproteins. Biochemistry 15:1808–1814
    [Google Scholar]
  8. Horisberger M., Rosset J., Bauer H. 1976; Localization of mannan at the surface of yeast protoplasts by scanning electron microscopy. Archives of Microbiology 109:9–14
    [Google Scholar]
  9. Kolarova N., Masler L., Sikl D. 1973; Cell wall glycoproteins of Candida albicans serotypes A and B. Biochimica et biophysica acta 328:221–227
    [Google Scholar]
  10. Korn E. D., Northcote D. H. 1960; Physical and chemical properties of polysaccharides and glycoproteins of the yeast cell wall. Biochemical Journal 75:12–17
    [Google Scholar]
  11. Lehle L., Tanner W. 1974; Membrane bound mannosyl transferases in yeast glycoprotein biosynthesis. Biochimica et biophysica acta 350:225–235
    [Google Scholar]
  12. Marchalonis J. J. 1969; An enzymic method for the trace iodination of immunoglobulins and other proteins. Biochemical Journal 113:299–305
    [Google Scholar]
  13. Marriott M. S. 1975a; Isolation and chemical characterization of plasma membranes from the yeast and mycelial forms of Candida albicans. Journal of General Microbiology 86:115–132
    [Google Scholar]
  14. Marriott M. S. 1975b; Enzymic activity of purified plasma membranes from the yeast and mycelial forms of Candida albicans. Journal of General Microbiology 89:345–352
    [Google Scholar]
  15. Morrison M. 1974; The determination of the exposed proteins on membranes by the use of lactoperoxidase. Methods in Enzymology 32B:103–109
    [Google Scholar]
  16. Nakajima T., Ballou C. E. 1974; Structure of the linkage region between the polysaccharide and protein parts of Saccharomyces cerevisiaemannan. Journal of Biological Chemistry 249:7685–7694
    [Google Scholar]
  17. Nakajima T., Ballou C. E. 1975; Yeast manno-protein biosynthesis; solubilization and selective assay of four mannosyl transferases. Proceedings of the National Academy of Sciences of the United States of America 72:3912–3916
    [Google Scholar]
  18. Phillips D. R., Morrison M. 1970; The arrangement of proteins in the human erythrocyte membrane. Biochemical and Biophysical Research Communications 40:284–289
    [Google Scholar]
  19. Phillips D. R., Morrison M. 1971; Exposed proteins of the intact human erythrocyte. Bio-chemistry 10:1766–1771
    [Google Scholar]
  20. Poduslo J. F., Greenberg C. S., Glick M. C. 1972; Proteins exposed on the surface of mammalian cell membranes. Biochemistry 11:2616–2621
    [Google Scholar]
  21. Salton M. R. J., Schor M. T., Ng M. H. 1972; Internal localization of Micrococcus lysodeikticus membrane ATPase by iodination with 125I. Biochimica et biophysica acta 290:408–413
    [Google Scholar]
  22. Schibeci A., Rattray J. B. M., Kidby D. K. 1973a; Isolation and identification of yeast plasma membrane. Biochimica et biophysica acta 311:15–25
    [Google Scholar]
  23. Schibeci A., Rattray J. B. M., Kidby D. K. 1973b; Electron microscope autoradiography of labelled yeast plasma membrane. Biochimica et biophysica acta 323:532–538
    [Google Scholar]
  24. Sentandreu R., Northcote D. H. 1969; The characterization of oligosaccharides attached to threonine and serine in a mannan-protein obtained from the cell wall of yeast. Carbohydrate Research 100:584–585
    [Google Scholar]
  25. Sharma C. B., Babczinski P., Lehle L., Tanner W. 1974; The role of dolichol monophosphate in glycoprotein biosynthesis in Saccharomyces cerevisiae. European Journal of Biochemistry 46:35–41
    [Google Scholar]
  26. Sikl D., Masler L., Bauer S. 1965; The polysaccharides of yeasts and yeast-like microorganisms I : the extracellular surface mannan of Candida albicans Berkhout. Chemicke zvesti 19:21–27
    [Google Scholar]
  27. Wallach D. F. H., LIM P. S. 1973; A critical evaluation of plasma membrane fractionation. Biochimica et biophysica acta 200:211–254
    [Google Scholar]
  28. Wilson C. 1959; Quantitative determination of sugars on paper chromatograms. Analytical Chemistry 31:1199–1201
    [Google Scholar]
  29. Wilson G., Rose S. P., Fox C. F. 1970; The effect of membrane lipid unsaturation on glycoside transport. Biochemical and Biophysical Research Communications 38:617–623
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-103-1-51
Loading
/content/journal/micro/10.1099/00221287-103-1-51
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error