Skip to main content
SpringerLink
Log in

Isolation and identification of a 92-kDa stress induced protein from Candida albicans

  • Published:
Mycopathologia Aims and scope Submit manuscript

Abstract

It was previously shown that the presence of estrogen enhances survival of Candida albicans under heat and oxidative stresses. A 92-kDa protein is inducible by heat shock and estrogen in C. albicans. Previous studies have described this protein as hsp90 because of its molecular size and heat inducibility as seen on electrophoretic gels and Western blots. In this study, ion exchange, hydroxy apatite and size exclusion chromatography were used to isolate a 92-kDa-protein band. The N-terminal sequence of isolated protein blotted onto a PVDF membrane was determined to be V-Q-S-?-V-L-G-F-P-R. This sequence is homologous to the N-terminal sequence of the MET6 gene product, cobalamin-independent methionine synthase, from Saccharomyces cerevisiae. The results of this study suggest that a cobalamin-independent methionine synthase homolog is inducible by heat and estrogen in C. albicans. This study also suggests that Candida hsp90 is more likely to exist as an 82-kDa protein as predicted by a previously described cDNA and not as a 92-kDa protein as reported in the literature.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Odds FC. Candida Species and Virulence. ASM News 1994; 60(6): 313–318.

    Google Scholar 

  2. Cutler J. Putative Virulence factors of Candida albicans. Annu. Rev. Microbiol. 1991; 45: 187–218.

    Google Scholar 

  3. White S, Larsen B. Candida albicans morphogenesis is influenced by estrogen. Cell Mol Life Sci 1997; 53(9): 744–749.

    Google Scholar 

  4. Gow NAR. Growth and Guidance of the Fungal Hypha. Microbiology 1994; 140: 3193.

    Google Scholar 

  5. Shah DT, Glover DD, Larsen B. In situ mycotoxin production by Candida albicans in women with vaginitis. Gynecol Obstet Invest 1995; 39(1): 67–69.

    Google Scholar 

  6. Gujjar PR, Finucane M, Larsen B. The effect of estradiol on Candida albicans growth. Ann Clin Lab Sci 1997; 27(2): 151–156.

    Google Scholar 

  7. O'Connor C, Essmann M, Larsen B. 17-beta-estradiol upregulates the stress response in Candida albicans: implications for microbial virulence. Infect Dis Obstet Gynecol 1998; 6(4): 176–181.

    Google Scholar 

  8. Mager WH, Ferreira PM. Stress response of yeast. Biochem J 1993; 290(Pt 1): 1–13.

    Google Scholar 

  9. Louvion JF, Abbas-Terki T, Picard D. Hsp90 Is Required for Pheromone Signaling in Yeast. Mol Biol Cell 1998; 9(11): 3071–3083.

    Google Scholar 

  10. Pratt WB. The role of the hsp90-based chaperone system in signal transduction by nuclear receptors and receptor signaling via MAP kinase. Annual Review of Pharmacology and Toxicology 1997; 37: 297–326.

    Google Scholar 

  11. Stancato LF, Silverstein AM, OwensGrillo JK, Chow YH, Jove R, Pratt WB. The hsp90-binding antibiotic geldanamycin decreases Raf levels and epidermal growth factor signaling without disrupting formation of signaling complexes or reducing the specific enzymatic activity of raf kinase. Journal of Biological Chemistry 1997; 272 (7): 4013–4020.

    Google Scholar 

  12. Nathan DF, Vos MH, Lindquist S. In vivo functions of the Saccharomyces cerevisiae Hsp90 chaperone. Proceedings of the National Academy of Sciences of the United States of America 1997; 94 (24): 12949–12956.

    Google Scholar 

  13. Swoboda RK, Bertram G, Budge S, Gooday GW, Gow NA, Brown AJ. Structure and regulation of the HSP90 gene from the pathogenic fungus Candida albicans [published erratum appears in Infect Immun 1996 Feb; 64(2): 680]. Infect Immun 1995; 63(11): 4506–4514.

    Google Scholar 

  14. Hodgetts S, Matthews R, Morrissey G, Mitsutake K, Piper P, Burnie J. Over-expression of Saccharomyces cerevisiae hsp90 enhances the virulence of this yeast in mice. FEMS Immunol Med Microbiol 1996; 16: 229–234.

    Google Scholar 

  15. Matthews R, Burnie J. The role of hsp90 in fungal infection. Immunol Today 1992; 13(9): 345–348.

    Google Scholar 

  16. Matthews RC, Burnie JP, Tabaqchali S. Isolation of immunodominant antigens from sera of patients with systemic candidiasis and characterization of serological response to Candida albicans. J Clin Microbiol 1987; 25(2): 230–237.

    Google Scholar 

  17. Matthews R, Burnie J. Cloning of a DNA sequence encoding a major fragment of the 47 kilodalton stress protein homologue of Candida albicans. FEMS Microbiol Lett 1989; 51(1): 25–30.

    Google Scholar 

  18. Matthews RC. HSP90, yeasts and Corynebacterium jeikeium. Epidemiol. Infect. 1991; 107: 273–283.

    Google Scholar 

  19. Srivastava PK. Purification of heat shock protein - Peptide complexes for use in vaccination against cancers and intracellular pathogens. Methods a Companion to Methods in Enzymology 1997; 12(2): 165–171.

    Google Scholar 

  20. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680–685.

    Google Scholar 

  21. Cleveland DW, Fischer SG, Kirschner MW, Laemmli UK. Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J Biol Chem 1977; 252(3): 1102–1106.

    Google Scholar 

  22. Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 1979; 76(9): 4350–4354.

    Google Scholar 

  23. Burnette WN. “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; 112(2): 195–203.

    Google Scholar 

  24. Strickler JE, Hunkapiller MW, Wilson KJ. Utility of the gasphase sequencer for both liquid-and solid-phase degradation of proteins and peptides at low picomole levels. Anal Biochem 1984; 140(2): 553–566.

    Google Scholar 

  25. Niall HD. Automated Edman degradation: the protein sequenator. Methods Enzymol 1973; 27: 942–1010.

    Google Scholar 

  26. Boucherie H, Dujardin G, Kermorgant M, Monribot C, Slonimski P, Perrot M. Two-dimensional protein map of Saccharomyces cerevisiae: construction of a gene-protein index. Yeast 1995; 11(7): 601–613.

    Google Scholar 

  27. Farrelly FW, Finkelstein DB. Complete sequence of the heat shock-inducible HSP90 gene of Saccharomyces cerevisiae. J Biol Chem 1984; 259(9): 5745–5751.

    Google Scholar 

  28. Zeuthen ML, Howard DH. Thermotolerance and the heatshock response in Candida albicans. J Gen Microbiol 1989; 135(Pt 9): 2509–2518.

    Google Scholar 

  29. Gething MJ, Sambrook J. Protein folding in the cell. Nature 1992; 355: 33–45.

    Google Scholar 

  30. Lindquist S, Craig EA. The heat-shock proteins. Annu Rev Genet 1988; 22: 631–677.

    Google Scholar 

  31. Balish E. Methionine biosynthesis and S-adenosylmethionine degradation during an induced morphogenesis of Candida albicans. Can J Microbiol 1973; 19(7): 847–853.

    Google Scholar 

  32. Godon C, Lagniel G, Lee J, Buhler JM, Kieffer S, Perrot M, et al. The H2O2 stimulon in Saccharomyces cerevisiae. J Biol Chem 1998; 273(34): 22480–22489.

    Google Scholar 

  33. Csermely P, Schnaider T, Soti C, Prohaszka Z, Nardai G. The 90-kDa molecular chaperone family: structure, function, and clinical applications. A comprehensive review. Pharmacol Ther 1998; 79(2): 129–168.

    Google Scholar 

  34. Szyszka R, Kramer G, Hardesty B. The phosphorylation state of the reticulocyte 90-kDa heat shock protein affects its ability 20 to increase phosphorylation of peptide initiation factor 2 alpha subunit by the heme-sensitive kinase. Biochemistry 1989; 28(4): 1435–1438.

    Google Scholar 

  35. Palmquist K, Riis B, Nilsson A, Nygard O. Interaction of the calcium and calmodulin regulated eEF-2 kinase with heat shock protein 90. FEBS Lett 1994;3 49(2): 239–242.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Biochemistry, Des Moines University–Osteopathic Medical Center, Des Moines, IA, 50312, USA

    Edward T. Burt

  2. Department of Biology, Drake University, Des Moines, IA, 50311, USA

    Christopher O'Connor

  3. Office of University Research, Des Moines University-Osteopathic Medical Center, Des Moines, IA, 50312, USA

    Bryan Larsen

Authors
  1. Edward T. Burt
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Christopher O'Connor
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Bryan Larsen
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Burt, E.T., O'Connor, C. & Larsen, B. Isolation and identification of a 92-kDa stress induced protein from Candida albicans . Mycopathologia 147, 13–20 (1999). https://doi.org/10.1023/A:1007036518330

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1007036518330

Search

Navigation