Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1986 Sep;53(3):565–572. doi: 10.1128/iai.53.3.565-572.1986

Analysis of cell wall extracts of Candida albicans by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot techniques.

J Ponton, J M Jones
PMCID: PMC260828  PMID: 3527986

Abstract

Cell walls of intact yeast- and mycelial-phase Candida albicans B311 were extracted with different compounds: dithiothreitol, dithiothreitol with protease, dithiothreitol with lyticase, and dithiothreitol with protease followed by beta-glucuronidase with chitinase. Extracts were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot techniques. Dithiothreitol extracts contained the most satisfactory array of components for study. Analysis of these extracts demonstrated that the outer cell wall layers of Candida blastoconidia and germ tubes contained a complex array of polysaccharides, glycoproteins, and proteins. The proteins contributed to a latticework stabilized by covalent bonds that was important in determining the porosity of the outer cell wall layers. When equivalent weights were analyzed, mycelial-phase extract contained a more varied array of proteins than did yeast-phase extract. Only a portion of proteins in mycelial-phase extract elicited antibody responses in hyperimmunized rabbits or infected humans. A polysaccharide-rich, high-molecular-weight component (migrating at a position that would correspond to proteins having molecular weights of 235,000 to 250,000) and a protein component (molecular weight, 19,000) were readily demonstrable in the mycelial-phase extract but could not be identified in the yeast-phase extract.

Full text

PDF
565

Images in this article

567Image
on p.567
567Image
on p.567
568Image
on p.568
569Image
on p.569
569Image
on p.569

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bruneau S. M., Guinet R. M. Characterization of Candida antigens by crossed-immunoaffinoelectrophoresis. Sabouraudia. 1985 Apr;23(2):107–118. [PubMed] [Google Scholar]
  2. Cassone A., Mattia E., Boldrini L. Agglutination of blastospores of Candida albicans by concanavalin A and its relationship with the distribution of mannan polymers and the ultrastructure of the cell wall. J Gen Microbiol. 1978 Apr;105(2):263–273. doi: 10.1099/00221287-105-2-263. [DOI] [PubMed] [Google Scholar]
  3. Cassone A., Simonetti N., Strippoli V. Ultrastructural changes in the wall during germ-tube formation from blastospores of Candida albicans. J Gen Microbiol. 1973 Aug;77(2):417–426. doi: 10.1099/00221287-77-2-417. [DOI] [PubMed] [Google Scholar]
  4. Centeno A., Davis C. P., Cohen M. S., Warren M. M. Modulation of Candida albicans attachment to human epithelial cells by bacteria and carbohydrates. Infect Immun. 1983 Mar;39(3):1354–1360. doi: 10.1128/iai.39.3.1354-1360.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. Chattaway F. W., Shenolikar S., O'Reilly J., Barlow A. J. Changes in the cell surface of the dimorphic forms of Candida albicans by treatment with hydrolytic enzymes. J Gen Microbiol. 1976 Aug;96(2):335–347. doi: 10.1099/00221287-95-2-335. [DOI] [PubMed] [Google Scholar]
  7. Elango N., Correa J. U., Cabib E. Secretory character of yeast chitinase. J Biol Chem. 1982 Feb 10;257(3):1398–1400. [PubMed] [Google Scholar]
  8. 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]
  9. Greenfield R. A., Stephens J. L., Bussey M. J., Jones J. M. Quantitation of antibody to Candida mannan by enzyme-linked immunosorbent assay. J Lab Clin Med. 1983 May;101(5):758–771. [PubMed] [Google Scholar]
  10. 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]
  11. Ho Y. M., Ng M. H., Huang C. T. Antibodies to germinating and yeast cells of Candida albicans in human and rabbit sera. J Clin Pathol. 1979 Apr;32(4):399–405. doi: 10.1136/jcp.32.4.399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Jones J. M. Quantitation of antibody against cell wall mannan and a major cytoplasmic antigen of Candida in rabbits, mice, and humans. Infect Immun. 1980 Oct;30(1):78–89. doi: 10.1128/iai.30.1.78-89.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kapitany R. A., Zebrowski E. J. A high resolution PAS stain for polyacrylamide gel electrophoresis. Anal Biochem. 1973 Dec;56(2):361–369. doi: 10.1016/0003-2697(73)90202-9. [DOI] [PubMed] [Google Scholar]
  14. Kimura L. H., Pearsall N. N. Adherence of Candida albicans to human buccal epithelial cells. Infect Immun. 1978 Jul;21(1):64–68. doi: 10.1128/iai.21.1.64-68.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kobayashi K., Suginaka H. Comparison of cell wall and membrane proteins from eight Candida species. Sabouraudia. 1984;22(4):341–344. [PubMed] [Google Scholar]
  16. 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]
  17. Lazdunski A., Murgier M., Lazdunski C. Evidence for an aminoendopeptidase localized near the cell surface of Escherichia coli. Regulation of synthesis by inorganic phosphate. Eur J Biochem. 1975 Dec 15;60(2):349–355. doi: 10.1111/j.1432-1033.1975.tb21009.x. [DOI] [PubMed] [Google Scholar]
  18. Lee J. C., King R. D. Characterization of Candida albicans adherence to human vaginal epithelial cells in vitro. Infect Immun. 1983 Sep;41(3):1024–1030. doi: 10.1128/iai.41.3.1024-1030.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. Lee K. L., Rega M. E., Watson R. R., Campbell C. C. Identification of yeast phase of pathogenic fungi by the specificity of their aminopeptidase(s). Sabouraudia. 1975 Jul;13(2):132–141. doi: 10.1080/00362177585190251. [DOI] [PubMed] [Google Scholar]
  21. Lehrer N., Segal E., Barr-Nea L. In vitro and in vivo adherence of Candida albicans to mucosal surfaces. Ann Microbiol (Paris) 1983 Sep-Oct;134B(2):293–306. doi: 10.1016/s0769-2609(83)80042-8. [DOI] [PubMed] [Google Scholar]
  22. Neu H. C., Heppel L. A. The release of enzymes from Escherichia coli by osmotic shock and during the formation of spheroplasts. J Biol Chem. 1965 Sep;240(9):3685–3692. [PubMed] [Google Scholar]
  23. 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]
  24. Poulter R., Jeffery K., Hubbard M. J., Shepherd M. G., Sullivan P. A. Parasexual genetic analysis of Candida albicans by spheroplast fusion. J Bacteriol. 1981 Jun;146(3):833–840. doi: 10.1128/jb.146.3.833-840.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Segal E., Soroka A., Lehrer N. Attachment of Candida to mammalian tissues--clinical and experimental studies. Zentralbl Bakteriol Mikrobiol Hyg A. 1984 Jul;257(2):257–265. [PubMed] [Google Scholar]
  26. Smail E. H., Jones J. M. Demonstration and solubilization of antigens expressed primarily on the surfaces of Candida albicans germ tubes. Infect Immun. 1984 Jul;45(1):74–81. doi: 10.1128/iai.45.1.74-81.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. 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]
  28. Sundstrom P. M., Kenny G. E. Characterization of antigens specific to the surface of germ tubes of Candida albicans by immunofluorescence. Infect Immun. 1984 Mar;43(3):850–855. doi: 10.1128/iai.43.3.850-855.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sundstrom P. M., Kenny G. E. Enzymatic release of germ tube-specific antigens from cell walls of Candida albicans. Infect Immun. 1985 Sep;49(3):609–614. doi: 10.1128/iai.49.3.609-614.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Torres-Bauzá L. J., Riggsby W. S. Protoplasts from yeast and mycelial forms of Candida albicans. J Gen Microbiol. 1980 Aug;119(2):341–349. doi: 10.1099/00221287-119-2-341. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. Zacharius R. M., Zell T. E., Morrison J. H., Woodlock J. J. Glycoprotein staining following electrophoresis on acrylamide gels. Anal Biochem. 1969 Jul;30(1):148–152. doi: 10.1016/0003-2697(69)90383-2. [DOI] [PubMed] [Google Scholar]
  33. 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]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES