Isolation and Chemical Characterization of Plasma Membranes from the Yeast and Mycelial Forms of Free

Abstract

SUMMARY: It has been possible to induce the yeast-mycelium transformation in by growth of the organism under completely defined conditions in batch culture. Protoplasts have been obtained from the two forms by using a lytic enzyme preparation from A plasma membrane fraction was prepared by osmotic lysis of these protoplasts and fractionated by using a combination of differential and discontinuous sucrose density-gradient flotation centrifugation. The purity of this fraction was determined by radioactive dansylation and iodination of plasma membranes of intact protoplasts followed by localization of the radioactivity upon fractionation. This procedure demonstrated less than 4% contamination of the plasma membrane fraction with other cell membranes. Chemical analysis of this fraction revealed that the major components were protein and lipid. Membranes from the yeast form contained (w/w): 50% protein, 45% lipid, 9% carbohydrate and 0·3% nucleic acid. Plasma membranes from the mycelial form contained significantly more carbohydrate and were found to be composed of (w/w): 43% protein, 31% lipid, 25% carbohydrate and 0·5% nucleic acid. Marked differences were also observed between the phospholipid, free and esterified sterols, and total fatty acids of membranes from the two forms of the organism.

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1975-01-01
2024-03-29
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References

  1. Anderson F. B., Millbank J. W. 1966; Protoplast formation and yeast cell wall structure. Biochemical Journal 99:682–687
    [Google Scholar]
  2. Baird J. K., Cunningham W. L. 1971; Formation of yeast protoplasts by using an enzyme preparation from Cytophaga. Biochemical Journal 125:32P–33P
    [Google Scholar]
  3. Barton D. H. R., Corrie J. E. T., Marshall P. J., Widdowson D. A. 1973; Biosynthesis of terpenes and steroids. VII. Unified scheme for the biosynthesis of ergosterol in Saccharomyces cerevisiae. Bioorganic Chemistry 2:363–373
    [Google Scholar]
  4. Bittman R., Fischkoff S. A. 1972; Fluorescence studies of the binding of the polyene antibiotics filipin III, amphotericin B, nystatin and lagosin to cholesterol. Proceedings of the National Academy of Sciences of the United States of America 693795–3799
    [Google Scholar]
  5. Boulton A. A., Eddy A. A. 1962; The properties of certain particles isolated from yeast protoplasts disrupted by osmotic shock. Biochemical Journal 82:16P–17P
    [Google Scholar]
  6. Breivik O. N., Owades J. L. 1957; A semi-micro method for the determination of the percentages of ergosterol and 24,28 dehydro-ergosterol in yeast fat. Journal of Agricultural and Food Chemistry 5:360–363
    [Google Scholar]
  7. Bretscher M. S. 1973a; Membrane structure: some general principles. Science; New York: 181622–629
    [Google Scholar]
  8. Bretscher M. S. 1973b; On labelling membranes. Nature New Biology 245:116–117
    [Google Scholar]
  9. Brown J. P., Perham R. N. 1973; A highly sensitive method for amino acid analysis by double-isotope-labelling technique using dansyl chloride. European Journal of Biochemistry 39:69–73
    [Google Scholar]
  10. 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]
  11. Chen P. S., Toribara T. Y., Warner H. 1956; Microdetermination of phosphorus. Analytical Chemistry 28:1756–1758
    [Google Scholar]
  12. Crane F. L., Mii S., Hauge J. G., Green D. E., Beinert H. 1956; On the mechanism of dehydrogenation of fatty acyl derivatives of coenzyme A. I. The general fatty acyl coenzyme A dehydrogenase. Journal of Biological Chemistry 218:701–716
    [Google Scholar]
  13. Dittmer J. C., Lester R. L. 1964; A simple, specific spray for the detection of phospholipids on thin layer chromatograms. Journal of Lipid Research 5:126–127
    [Google Scholar]
  14. Drabikowski W., Lagwinska E., Sarzala M. G. 1973; Filipin as a fluorescent probe for the location of cholesterol in the membranes of fragmented sarcoplasmic reticulum. Biochimica et biophysica acta 291:61–70
    [Google Scholar]
  15. Dubé J., Setterfield G., Kiss G., Lusena C. V. 1973; Fate of the plasma-membrane of Saccharomyces cerevisiae during cell rupture. Canadian Journal of Microbiology 19:285–290
    [Google Scholar]
  16. Elorza M. V., Munoz-Ruiz E., Villanueva J. R. 1966; Production of yeast cell wall lytic enzymes on a semi-defined medium by a Streptomyces. Nature; London: 210442–443
    [Google Scholar]
  17. Exley D. 1957; The determination of 10-100 ng quantities of hexosamine. Biochemical Journal 67:52–60
    [Google Scholar]
  18. Folch J., Lees M., Sloane-Stanley G. H. 1957; A Simple Method For The Isolation And Purification Of Total Lipids From Animal Tissues. Journal Of Biological Chemistry 226:497–509
    [Google Scholar]
  19. Garcia-Mendoza C., Villanueva J. R. 1962; Production of yeast protoplasts by an enzyme preparation of Streptomyces species. Nature; London: 1951326–1327
    [Google Scholar]
  20. Garcia-Mendoza C., Villanueva J. R. 1967; Preparation and composition of the protoplast membrane of Candida utilis. Biochimica et biophysica acta 136:189–195
    [Google Scholar]
  21. Ghuysen J.-M., Tipper D. J., Strominger J. L. 1966; Enzymes that degrade bacterial cell walls. In Methods in Enzymology 8 pp. 685–699 Neufeld E. F., Ginsburg K. Edited by New York and London: Academic Press;
    [Google Scholar]
  22. Green D. E., Mii S., Kohout P. M. 1955; Studies on the terminal electron transport system. I. Succinic dehydrogenase. Journal of Biological Chemistry 217:551–567
    [Google Scholar]
  23. Gull K., Moore P. M., Trinci A. P. J. 1972; Formation of protoplasts from Geotrichum lactis and use of the fluorescence to detect cell walls. Transactions of the British Mycological Society 59:79–85
    [Google Scholar]
  24. Heinen W., De Vries H. 1966; A combined micro and semi-micro colorimetric determination of long chain fatty acids from plant cutin. Archiv für Mikrobiologie 54:339–349
    [Google Scholar]
  25. James A. T., Martin A. J. P. 1952; Gas-liquid partition chromatography: the separation and microestimation of volatile fatty acids from formic to dodecanoic acid. Biochemical Journal 50:679–690
    [Google Scholar]
  26. Kassai M., Podleski T. R., Changeux J. P. 1970; Some structural properties of excitable membranes labelled by fluorescent probes. FEBS Letters 7:13–19
    [Google Scholar]
  27. Kobayashi G. S., Friedman L., Kofroth J. F. 1964; Some cytological and pathogenic properties of spheroplasts of Candida albicans. Journal of Bacteriology 88:795–801
    [Google Scholar]
  28. Lampen J. O. 1968; External enzymes of yeast: their nature and formation. Antonie van Leeuwenhoek 34:1–18
    [Google Scholar]
  29. Lampen J. O., Arnow P., Borowska Z., Laskin A. I. 1962; Location and role of sterol at nystatin binding sites. Journal of Bacteriology 84:1152–1160
    [Google Scholar]
  30. Longley R. P., Rose A. H., Knights B. A. 1968; Composition of the protoplast membrane of Saccharomyces cerevisiae. Biochemical Journal 108:401–412
    [Google Scholar]
  31. Lowry O. H., Rosebrough N. J., Farr A. L., Randall R. J. 1951; Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 193:265–275
    [Google Scholar]
  32. Mardon D. N., Balish E., Phillips A. W. 1969; Control of dimorphism in a biochemical variant of Candida albicans. Journal of Bacteriology 100:701–707
    [Google Scholar]
  33. Mardon D. N., Hurst S. K., Balish E. 1971; Germ tube production by Candida albicans in minimal culture media (liquid). Canadian Journal of Microbiology 17:851–856
    [Google Scholar]
  34. Matile P. 1970; Properties of the purified cytoplasmic membrane of yeast. Federation of European Biochemical Societies Symposium 20 pp. 39–49 Villanueva J. R., Ponz F. Edited by New York and London: Academic Press;
    [Google Scholar]
  35. Matile P., Moor H., Mühlethaler K. 1967; Isolation and properties of plasmalemma in yeast. Archiv für Mikrobiologie 58:201–211
    [Google Scholar]
  36. Nichols B. W., Moorhouse R. 1969; The separation, structure and metabolism of monogalactosyl diglyceride species in Chlorella vulgaris. Lipids 4:311–316
    [Google Scholar]
  37. Nickerson W. J., Mankowski Z. 1953; Role of nutrition in the maintenance of yeast shape in Candida. American Journal of Botany 40:584–592
    [Google Scholar]
  38. Nombela C., Uruburu F., Villanueva J. R. 1974; Studies on membranes isolated from extracts of Fusarium culmorum. Journal of General Microbiology 81:247–254
    [Google Scholar]
  39. Nurminen T., Oura E., Suomalainen H. 1970; The enzymic composition of the isolated cell wall and plasma-membrane of baker’s yeast. Biochemical Journal 116:61–69
    [Google Scholar]
  40. Rondle C. M., Morgan W. T. J. 1955; The determination of glucosamine and galactosamine. Biochemical Journal 61:586–589
    [Google Scholar]
  41. 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]
  42. 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]
  43. 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]
  44. Skidmore W. D., Entenman C. 1962; Two dimensional thin-layer chromatography of rat liver phosphatides. Journal of Lipid Research 3:471–475
    [Google Scholar]
  45. Skipski V. P., Peterson R. F., Barclay M. 1964; Quantitative analysis of phospholipids by thin-layer chromatography. Biochemical Journal 90:374–378
    [Google Scholar]
  46. Starr P. R., Parks L. W. 1972; Transmethylation of sterols in aerobically adapting Saccharomyces cerevisiae. Journal of Bacteriology 109:236–242
    [Google Scholar]
  47. Sweeley C. C., Bentley R., Makita M., Wells W. W. 1963; Gas-liquid chromatography of trimethyl-silyl derivatives of sugars and related substances. Journal of the American Chemical Society 85:2497–2507
    [Google Scholar]
  48. Tkacz J. S., Cybulska E. B., Lampen J. O. 1971; Specific staining of wall mannan in yeast cells with fluorescein conjugated Concanavalin A. Journal of Bacteriology 105:1–5
    [Google Scholar]
  49. Trevelyan W. E., Harrison J. S. 1952; Fractionation and microdetermination of cell carbohydrates. Biochemical Journal 50:298–305
    [Google Scholar]
  50. Vandenheuval F. A., Court A. S. 1968; Reference high efficiency non polar packed columns for the gas-liquid chromatography of nanogram amounts of steroids. I. Retention time data. Journal of Chromatography 38:439–459
    [Google Scholar]
  51. Van Handel E., Zilversmit D. B. 1957; Micromethod for the determination of serum triglycerides. Journal of Laboratory and Clinical Medicine 50:152–157
    [Google Scholar]
  52. Widra A. 1964; The influence of the dextrose/phosphate ratio, nitrogen source and various cations on filamentation in Candida albicans. Mycopathologia et mycologia applicata 23:197–202
    [Google Scholar]
  53. Wood P., Imaichi K., Knowles J., Michaels C., Kinsell L. 1964; The lipid composition of human plasma chylomicrons. Journal of Lipid Research 5:225–231
    [Google Scholar]
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