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. 1991 Jul;59(7):2324–2332. doi: 10.1128/iai.59.7.2324-2332.1991

Characterization of cell wall proteins of yeast and hydrophobic mycelial cells of Candida albicans.

J L Lopez-Ribot 1, M Casanova 1, J P Martinez 1, R Sentandreu 1
PMCID: PMC258014  PMID: 2050401

Abstract

Cell surface hydrophobicity (CSH) of blastoconidia and blastoconidia bearing germ tubes of Candida albicans ATCC 26555 was monitored by assessing attachment of polystyrene microspheres to the cell surface, and we found that mature hyphae were significantly hydrophobic. Treatment of intact cells with low concentrations of beta-glucanase (Zymolyase 20T) or proteases abolished or significantly reduced attachment of latex beads to hyphae. This effect paralleled an obvious reduction in CSH of the entire cell population, as measured by an aqueous-hydrocarbon biphasic partitioning assay. Analysis of the cell wall material released by Zymolyase and adsorbed on polystyrene microspheres indicated that germ tube-specific cell wall proteins and mannoproteins with apparent molecular masses of 20 to 67 kDa may be responsible for the hydrophobicity of hyphae. Zymolyase released from blastoconidia cell walls a different set of proteins and mannoproteins that were able to adsorb to polystyrene microbeads. Such molecular species might in turn be responsible for the CSH exhibited by blastoconidium populations as determined by the biphasic partitioning assay, although these probably hydrophobic components can be masked on the surface of blastoconidia, as the latter had no or very few latex microspheres attached to their surfaces. Treatment of cells of both C. albicans morphologies with 2-mercaptoethanol released qualitatively distinct species of polystyrene-adsorbed proteins and mannoproteins from yeast and mycelial cells. These observations suggested that hydrophobic proteins and mannoproteins that could be associated with CSH are bound to the cell wall structure through diverse types of linkages.

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Selected References

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  1. Beachey E. H. Bacterial adherence: adhesin-receptor interactions mediating the attachment of bacteria to mucosal surface. J Infect Dis. 1981 Mar;143(3):325–345. doi: 10.1093/infdis/143.3.325. [DOI] [PubMed] [Google Scholar]
  2. Borg M., Rüchel R. Expression of extracellular acid proteinase by proteolytic Candida spp. during experimental infection of oral mucosa. Infect Immun. 1988 Mar;56(3):626–631. doi: 10.1128/iai.56.3.626-631.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bouali A., Robert R., Tronchin G., Senet J. M. Characterization of binding of human fibrinogen to the surface of germ-tubes and mycelium of candida albicans. J Gen Microbiol. 1987 Mar;133(3):545–551. doi: 10.1099/00221287-133-3-545. [DOI] [PubMed] [Google Scholar]
  4. Bouchara J. P., Tronchin G., Annaix V., Robert R., Senet J. M. Laminin receptors on Candida albicans germ tubes. Infect Immun. 1990 Jan;58(1):48–54. doi: 10.1128/iai.58.1.48-54.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brawner D. L., Cutler J. E. Ultrastructural and biochemical studies of two dynamically expressed cell surface determinants on Candida albicans. Infect Immun. 1986 Jan;51(1):327–336. doi: 10.1128/iai.51.1.327-336.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Burnette W. N. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. doi: 10.1016/0003-2697(81)90281-5. [DOI] [PubMed] [Google Scholar]
  7. Calderone R. A., Linehan L., Wadsworth E., Sandberg A. L. Identification of C3d receptors on Candida albicans. Infect Immun. 1988 Jan;56(1):252–258. doi: 10.1128/iai.56.1.252-258.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Casanova M., Gil M. L., Cardeñoso L., Martinez J. P., Sentandreu R. Identification of wall-specific antigens synthesized during germ tube formation by Candida albicans. Infect Immun. 1989 Jan;57(1):262–271. doi: 10.1128/iai.57.1.262-271.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Casanova M., Martínez J. P., Chaffin W. L. Fab fragments from a monoclonal antibody against a germ tube mannoprotein block the yeast-to-mycelium transition in Candida albicans. Infect Immun. 1990 Nov;58(11):3810–3812. doi: 10.1128/iai.58.11.3810-3812.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cassone A. Cell wall of Candida albicans: its functions and its impact on the host. Curr Top Med Mycol. 1989;3:248–314. doi: 10.1007/978-1-4612-3624-5_10. [DOI] [PubMed] [Google Scholar]
  11. Chaffin W. L., Skudlarek J., Morrow K. J. Variable expression of a surface determinant during proliferation of Candida albicans. Infect Immun. 1988 Feb;56(2):302–309. doi: 10.1128/iai.56.2.302-309.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Chaffin W. L., Stocco D. M. Cell wall proteins of Candida albicans. Can J Microbiol. 1983 Oct;29(10):1438–1444. doi: 10.1139/m83-220. [DOI] [PubMed] [Google Scholar]
  13. Critchley I. A., Douglas L. J. Isolation and partial characterization of an adhesin from Candida albicans. J Gen Microbiol. 1987 Mar;133(3):629–636. doi: 10.1099/00221287-133-3-629. [DOI] [PubMed] [Google Scholar]
  14. Douglas L. J. Adhesion of Candida species to epithelial surfaces. Crit Rev Microbiol. 1987;15(1):27–43. doi: 10.3109/10408418709104446. [DOI] [PubMed] [Google Scholar]
  15. Edison A. M., Manning-Zweerink M. Comparison of the extracellular proteinase activity produced by a low-virulence mutant of Candida albicans and its wild-type parent. Infect Immun. 1988 May;56(5):1388–1390. doi: 10.1128/iai.56.5.1388-1390.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Elorza M. V., Murgui A., Sentandreu R. Dimorphism in Candida albicans: contribution of mannoproteins to the architecture of yeast and mycelial cell walls. J Gen Microbiol. 1985 Sep;131(9):2209–2216. doi: 10.1099/00221287-131-9-2209. [DOI] [PubMed] [Google Scholar]
  17. Elorza M. V., Rico H., Gozalbo D., Sentandreu R. Cell wall composition and protoplast regeneration in Candida albicans. Antonie Van Leeuwenhoek. 1983 Nov;49(4-5):457–469. doi: 10.1007/BF00399324. [DOI] [PubMed] [Google Scholar]
  18. Elorza V., Mormeneo S., Garcia de la Cruz F., Gimeno C., Sentandreu R. Evidence for the formation of covalent bonds between macromolecules in the domain of the wall of Candida albicans mycelial cells. Biochem Biophys Res Commun. 1989 Aug 15;162(3):1118–1125. doi: 10.1016/0006-291x(89)90789-4. [DOI] [PubMed] [Google Scholar]
  19. Gilmore B. J., Retsinas E. M., Lorenz J. S., Hostetter M. K. An iC3b receptor on Candida albicans: structure, function, and correlates for pathogenicity. J Infect Dis. 1988 Jan;157(1):38–46. doi: 10.1093/infdis/157.1.38. [DOI] [PubMed] [Google Scholar]
  20. Hawkes R. Identification of concanavalin A-binding proteins after sodium dodecyl sulfate--gel electrophoresis and protein blotting. Anal Biochem. 1982 Jun;123(1):143–146. doi: 10.1016/0003-2697(82)90634-0. [DOI] [PubMed] [Google Scholar]
  21. Hazen B. W., Hazen K. C. Dynamic expression of cell surface hydrophobicity during initial yeast cell growth and before germ tube formation of Candida albicans. Infect Immun. 1988 Sep;56(9):2521–2525. doi: 10.1128/iai.56.9.2521-2525.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hazen K. C., Lay J. G., Hazen B. W., Fu R. C., Murthy S. Partial biochemical characterization of cell surface hydrophobicity and hydrophilicity of Candida albicans. Infect Immun. 1990 Nov;58(11):3469–3476. doi: 10.1128/iai.58.11.3469-3476.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Hazen K. C., LeMelle W. G. Improved assay for surface hydrophobic avidity of Candida albicans cells. Appl Environ Microbiol. 1990 Jun;56(6):1974–1976. doi: 10.1128/aem.56.6.1974-1976.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Hazen K. C. Participation of yeast cell surface hydrophobicity in adherence of Candida albicans to human epithelial cells. Infect Immun. 1989 Jul;57(7):1894–1900. doi: 10.1128/iai.57.7.1894-1900.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Hazen K. C., Plotkin B. J., Klimas D. M. Influence of growth conditions on cell surface hydrophobicity of Candida albicans and Candida glabrata. Infect Immun. 1986 Oct;54(1):269–271. doi: 10.1128/iai.54.1.269-271.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Kennedy M. J., Sandin R. L. Influence of growth conditions on Candida albicans adhesion, hydrophobicity and cell wall ultrastructure. J Med Vet Mycol. 1988 Apr;26(2):79–92. [PubMed] [Google Scholar]
  27. Kimura L. H., Pearsall N. N. Relationship between germination of Candida albicans and increased adherence to human buccal epithelial cells. Infect Immun. 1980 May;28(2):464–468. doi: 10.1128/iai.28.2.464-468.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Klotz S. A., Drutz D. J., Zajic J. E. Factors governing adherence of Candida species to plastic surfaces. Infect Immun. 1985 Oct;50(1):97–101. doi: 10.1128/iai.50.1.97-101.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  30. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  31. Lee K. L., Buckley H. R., Campbell C. C. An amino acid liquid synthetic medium for the development of mycelial and yeast forms of Candida Albicans. Sabouraudia. 1975 Jul;13(2):148–153. doi: 10.1080/00362177585190271. [DOI] [PubMed] [Google Scholar]
  32. Macura A. B. Hydrophobicity of Candida albicans related to their adherence to mucosal epithelial cells. Zentralbl Bakteriol Mikrobiol Hyg A. 1987 Oct;266(3-4):491–496. doi: 10.1016/s0176-6724(87)80231-6. [DOI] [PubMed] [Google Scholar]
  33. Millette C. F., Scott B. K. Identification of spermatogenic cell plasma membrane glycoproteins by two-dimensional electrophoresis and lectin blotting. J Cell Sci. 1984 Jan;65:233–248. doi: 10.1242/jcs.65.1.233. [DOI] [PubMed] [Google Scholar]
  34. Ponton J., Jones J. M. Analysis of cell wall extracts of Candida albicans by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot techniques. Infect Immun. 1986 Sep;53(3):565–572. doi: 10.1128/iai.53.3.565-572.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Poulain D., Hopwood V., Vernes A. Antigenic variability of Candida albicans. Crit Rev Microbiol. 1985;12(3):223–270. doi: 10.3109/10408418509104430. [DOI] [PubMed] [Google Scholar]
  36. Rotrosen D., Calderone R. A., Edwards J. E., Jr Adherence of Candida species to host tissues and plastic surfaces. Rev Infect Dis. 1986 Jan-Feb;8(1):73–85. doi: 10.1093/clinids/8.1.73. [DOI] [PubMed] [Google Scholar]
  37. Saxena A., Calderone R. Purification and characterization of the extracellular C3d-binding protein of Candida albicans. Infect Immun. 1990 Feb;58(2):309–314. doi: 10.1128/iai.58.2.309-314.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Skerl K. G., Calderone R. A., Segal E., Sreevalsan T., Scheld W. M. In vitro binding of Candida albicans yeast cells to human fibronectin. Can J Microbiol. 1984 Feb;30(2):221–227. doi: 10.1139/m84-033. [DOI] [PubMed] [Google Scholar]
  39. Sobel J. D., Muller G., Buckley H. R. Critical role of germ tube formation in the pathogenesis of candidal vaginitis. Infect Immun. 1984 Jun;44(3):576–580. doi: 10.1128/iai.44.3.576-580.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Sundstrom P. M., Nichols E. J., Kenny G. E. Antigenic differences between mannoproteins of germ tubes and blastospores of Candida albicans. Infect Immun. 1987 Mar;55(3):616–620. doi: 10.1128/iai.55.3.616-620.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Sundstrom P. M., Tam M. R., Nichols E. J., Kenny G. E. Antigenic differences in the surface mannoproteins of Candida albicans as revealed by monoclonal antibodies. Infect Immun. 1988 Mar;56(3):601–606. doi: 10.1128/iai.56.3.601-606.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Tronchin G., Bouchara J. P., Robert R. Dynamic changes of the cell wall surface of Candida albicans associated with germination and adherence. Eur J Cell Biol. 1989 Dec;50(2):285–290. [PubMed] [Google Scholar]
  44. Tronchin G., Bouchara J. P., Robert R., Senet J. M. Adherence of Candida albicans germ tubes to plastic: ultrastructural and molecular studies of fibrillar adhesins. Infect Immun. 1988 Aug;56(8):1987–1993. doi: 10.1128/iai.56.8.1987-1993.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Tronchin G., Robert R., Bouali A., Senet J. M. Immunocytochemical localization of in vitro binding of human fibrinogen to Candida albicans germ tube and mycelium. Ann Inst Pasteur Microbiol. 1987 Mar-Apr;138(2):177–187. doi: 10.1016/0769-2609(87)90194-3. [DOI] [PubMed] [Google Scholar]
  46. Zlotnik H., Fernandez M. P., Bowers B., Cabib E. Saccharomyces cerevisiae mannoproteins form an external cell wall layer that determines wall porosity. J Bacteriol. 1984 Sep;159(3):1018–1026. doi: 10.1128/jb.159.3.1018-1026.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]

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