1887

Abstract

Cell surface hydrophobicity (CSH) of grown both in complex medium (yeast extract/peptone/dextrose; YPD) and minimal (Vogel's N) medium was monitored by assessing attachment of polystyrene microspheres to the cell surface. It was found that mature mycelium was hydrophobic. Treatment of intact mycelium with β-mercaptoethanol (βME) abolished binding of the microspheres to hyphal elements, and coating of the microspheres with βME extracts from mycelium inhibited their attachment to intact mycelial cells. mycelium was tagged with biotin and treated with βME. The βME extracts were analysed by SDS-PAGE and Western blotting with both peroxidase-conjugated-ExtrAvidin and concanavalin A (ConA). This procedure allowed identification of cell wall surface proteins and glycoproteins. Rabbit polyclonal antisera were raised against βME extracts obtained from cells grown in YPD and Vogel's N media. These antisera defined some major cell-wall-bound antigens. SDS-PAGE and Western blotting analysis of the cell wall material released by βME and adsorbed on polystyrene microspheres revealed about 19 protein species with apparent molecular masses ranging from 20 to 70 kDa, and two high-molecular-mass glycoproteins of 115 and 210 kDa. Treatment of cells grown in YPD, but not those grown in Vogel's N medium, with βME released a 55 kDa polypeptide able to adsorb to polystyrene microspheres that was detectable with the antisera. The ability to bind to polystyrene particles exhibited by several protein and glycoprotein species released by βME treatment suggested that these cell wall moieties possess exposed hydrophobic domains that could be responsible for the CSH of mycelium.

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1996-07-01
2021-10-23
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References

  1. Annaix V., Bouchara J. P., Larcher G., Chabasse D., Tronchin G. 1992; Specific binding of fibrinogen fragment D to Aspergillus fumigatus conidia. Infect Immun 60:1747–1755
    [Google Scholar]
  2. Antley P.P., Hazen K. C. 1988; Role of yeast cell growth temperature on Candida albicans virulence in mice. Infect Immun 56:2884–2890
    [Google Scholar]
  3. Beachey E.H. 1981; Bacterial adherence : adhesin-receptor interactions mediating the attachment of bacteria to mucosal surfaces. J Infect Dis 143:325–345
    [Google Scholar]
  4. Bodey G.P., Vartivarían S. 1989; Aspergillosis. Eur J Clin Microbiol Infect Dis 8:413–437
    [Google Scholar]
  5. Bouchara J. P., Tronchin G., Larcher G., Chabasse D. 1995; The search for virulence determinants in Aspergillus fumigatus. Trends Microbiol 3:327–330
    [Google Scholar]
  6. Calderone R.A., Braun P. C. 1991; Adherence and receptor relationships in Candida albicans. Microbiol Rev 55:1–20
    [Google Scholar]
  7. Casanova M., Chaffin W. L. 1991; Cell wall glycoproteins of Candida albicans as released by different methods. J Gen Microbiol 137:1045–1051
    [Google Scholar]
  8. Casanova M., Gil M. L., Cardeñoso L., Martínez J. P., Sentandreu R. 1989; Identification of wall-specific antigens synthesized during germ tube formation by Candida albicans. Infect Immun 57:262–271
    [Google Scholar]
  9. Casanova M., López-Ribot J. L., Martínez J. P., Sentandreu R. 1992a; Characterization of cell wall proteins from yeast and mycelial cells of Candida albicans by labelling with biotin: comparison with other techniques. Infect Immun 60:4898–4906
    [Google Scholar]
  10. Casanova M., Lopez-Ribot J. L., Monteagudo C., LLombart-Bosch A., Sentandreu R., Martinez J. P. 1992b; Identification of a 58-kilodalton cell surface fibrinogen-binding mannoprotein from Candida albicans. Infect Immun 60:4221–4229
    [Google Scholar]
  11. Chaffin W.L., Stocco D. M. 1983; Cell wall proteins of Candida albicans. Can J Microbiol 29:1438–1444
    [Google Scholar]
  12. Cohen J. 1991; Clinical manifestations and management of aspergillosis in the compromised patient. In Fungal Infection in the Compromised Patient, 2nd edn. pp. 117–152 Edited by Warnock D. W., Richardson M. D. Chichester: John Wiley;
    [Google Scholar]
  13. Coulot P., Bouchara J. P., Renier G., Annaix V., Planchenault C., Tronchin G., Chabasse D. 1994; Specific interaction of Aspergillus fumigatus with fibrinogen and its role in cell adhesion. Infect Immun 62:2169–2177
    [Google Scholar]
  14. Diamond R.D. 1988; Fungal surfaces: effects of interactions with phagocytic cells. Rev Infect Dis 10:S428–S431
    [Google Scholar]
  15. Drouhet E., Camay L., Segretain G. 1972; Valeur de l’immunoprecipitation et de l’immunofluorescence indirecte dans les aspergilloses broncho-pulmonaires. Ann Inst Pasteur 123:379–395
    [Google Scholar]
  16. Dubois M., Gilles K. A., Hamilton J. K., Rebers P. A., Smith F. 1956; Colorimetric method for determination of sugars and related substances. Anal Chem 28:350–356
    [Google Scholar]
  17. Glee P. M., Sundstrom P., Hazen K. C. 1995; Expression of surface hydrophobic proteins by Candida albicans in vivo. Infect Immun 63:1373–1379
    [Google Scholar]
  18. Goldstein I.J., Hayes C. E. 1978; The lectins: carbohydrate- binding proteins of plants and animals. In Advances in Carbohydrate Chemistry and Biochemistry pp. 35127–340 Edited by Tipson R. S., Horton D. New York: Academic Press;
    [Google Scholar]
  19. Hazen K.C. 1990; Cell surface hydrophobicity of medically important fungi, especially Candida species. In Microbial Cell Surface Hydrophobicity pp. 249–295 Edited by Doyle R. J., Rosenberg M. . Washington, DC: American Society for Microbiology;
    [Google Scholar]
  20. Hazen K.C., Hazen B. W. 1987; A polystyrene microsphere assay for detecting surface hydrophobicity variations within Candida albicans populations. J Microbiol Methods 6:289–299
    [Google Scholar]
  21. Hazen K. C., Lay J. G., Hazen B. W., Fu R. C., Murthy S. 1990; Partial biochemical characterization of cell surface hydrophobicity and hydrophilicity of Candida albicans. Infect Immun 58:3469–3476
    [Google Scholar]
  22. Hearn V.M. 1984; Surface antigens of intact Aspergillus fumigatus mycelium: their localization using radiolabelled protein A as marker. J Gen Microbiol 130:907–917
    [Google Scholar]
  23. Hearn V.M. 1992; Antigenicity of Aspergillus species. J Med Vet Mycol 30:11–25
    [Google Scholar]
  24. Hearn V.M., Sietsma J. H. 1994; Chemical and immunological analysis of the Aspergillus fumigatus cell wall. Microbiology 140:789–795
    [Google Scholar]
  25. Hearn V. M., Wilson E. V., Latgé J.-P., Mackenzie D. W. R. 1990; Immunochemical studies of Aspergillus fumigatus mycelial antigens by polyacrylamide gel electrophoresis and Western blotting techniques. J Gen Microbiol 136:1525–1535
    [Google Scholar]
  26. Hearn V. M., Latgé J. P., Prévost M. C. 1991; Immuno- localization of Aspergillus fumigatus mycelial antigens. J Med Vet Mycol 29:73–81
    [Google Scholar]
  27. Kwon-Chung K.J., Bennett J. E. 1991; Aspergillosis.. In Medical Mycology pp. 201–247 Edited by Kwon-Chung K. J., Bennett J. E. Philadelphia: Lea and Febiger;
    [Google Scholar]
  28. Laemmli U.K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
    [Google Scholar]
  29. Latgé J.-P., Kobayashi H., Debeaupuis J. P., Diaquin M., Sarfati J., Wieruszeski J. M., Parra E., Bouchara J. P., Fournet B. 1994; Chemical and immunological characterization of the extracellular galactomannan of Aspergillus fumigatus. Infect Immun 62:5424–5433
    [Google Scholar]
  30. López-Ribot J. L., Casanova M., Martínez J.P., Sentandreu R. 1991; Characterization of cell wall proteins of yeast and hydrophobic mycelial cells of Candida albicans. Infect Immun 59:2324–2332
    [Google Scholar]
  31. López-Ribotones J.L., Gozalbo D., Sepúlveda P., Casanova M., Martínez J.P. 1995; Preliminary characterization of the material released to the culture medium by Candida albicans yeast and mycelial cells. Antonie Leeuwenhoek 68:195–201
    [Google Scholar]
  32. Martínez J.P., Casanova M., Gil M. L., Sentandreu R. 1991; Analysis of the polypeptide composition of the cell walls of Neurospora crassa. Similarities with the proteinaceous material secreted by the slime variant. Mycol Res 95:315–319
    [Google Scholar]
  33. Morrissey J.H. 1981; Silver stain for protein in polyacrylamide gels : a modified procedure with enhanced uniform sensitivity. Anal Biochem 117:307–310
    [Google Scholar]
  34. Parta M., Chang Y., Rulong S., Pinto-DaSilva P., Kwong- Chung K. J. 1994; HYP I, a hydrophobin gene from Aspergillus fumigatus, complements the rodletless phenotype in Aspergillus nidulans. Infect Immun 62:4389–4395
    [Google Scholar]
  35. Peñalver M.C., O'Connor J. E., Martínez J. P., Gil M. L. 1996; Binding of human fibronectin to Aspergillus fumigatus conidia. Infect Immun 64:1146–1153
    [Google Scholar]
  36. Ponton J., Jones J. M. 1986; Analysis of cell wall extracts of Candida albicans by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot techniques. Infect Immun 53:565–572
    [Google Scholar]
  37. Reijula K. E., Kurup V. P., Kumar A., Fink J. N. 1992; Monoclonal antibodies bind identically to both spores and hyphae of Aspergillus fumigatus. Clin Exp Allergy 22:547–553
    [Google Scholar]
  38. Rosenberg M., Gutnick D., Rosenberg E. 1980; Adherence of bacteria to hydrocarbons: a simple method for measuring cell- surface hydrophobicity. FEMS Microbiol Lett 9:29–33
    [Google Scholar]
  39. Rotrosen D., Calderone R. A., Edwards J. E. 1986; Adherence of Candida species to host tissues and plastic surfaces. Rev Infect Dis 8:73–85
    [Google Scholar]
  40. Silva T.M.J., Glee P. M., Hazen K. C. 1995; Influence of cell surface hydrophobicity on attachment of Candida albicans to extracellular matrix proteins. J Med Vet Mycol 33:117–122
    [Google Scholar]
  41. Ste-Marie L., Senechal S., Boushira M., Garzón S., Strykowski H., Pedneault L., de Repentigny L. 1990; Production and characterization of monoclonal antibodies to cell wall antigens of Aspergillus fumigatus. Infect Immun 58:2105–2114
    [Google Scholar]
  42. Sturtevant J.E., Latgé J.P. 1992; Interactions between conidia of Aspergillus fumigatus and human complement C3. Infect Immun 60:1913–1918
    [Google Scholar]
  43. Stynen D., Sarfati J., Goris A., Prévost M. C., Lesourd M., Kamphuis H., Darras V., Latgé J.-P. 1992; Rat monoclonal antibodies against Aspergillus galactomannan. Infect Immun 60:2237–2245
    [Google Scholar]
  44. Thau N., Monod M., Crestani B., Rolland C., Tronchin G., Latgé J.-P., Paris S. 1994; rodletless mutants of Aspergillus fumigatus. Infect Immun 62:4380–4388
    [Google Scholar]
  45. Tronchin G., Bouchara J. P., Larcher G., Lissitsky J. C., Chabasse D. 1993; Interaction between Aspergillus fumigatus and basement membrane laminin: binding and substrate degradation. Biol Cell 77:201–208
    [Google Scholar]
  46. Wilson E.V., Hearn V. M. 1982; A comparison of surface and cytoplasmic antigens of Aspergillus fumigatus in an enzyme-linked mmunosorbent assay (ELISA). Mykosen 25:653–661
    [Google Scholar]
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