1887

Abstract

Summary: , the most common fungal pathogen of humans, possesses an oestrogen (estrogen)-binding protein (EBP) that binds oestrogens with high affinity and specificity. The gene that encodes the EBP () has been cloned and sequenced and shown to be structurally related to the old yellow enzyme from . Here, we report the purification and the subcellular localization of the EBP from Using ion-exchange chromatography and an oestradiol affinity column, the EBP was purified from a strain of (strain 422) which was selected because it constitutively expressed elevated levels of the binding protein. The purified protein displayed a subunit molecular mass of approximately 46 kDa when examined by denaturing gel electrophoresis, which is consistent with the size estimated from the sequence of the cloned gene. An immunoaffinity column, prepared using a polyclonal antisera generated against EBP, depleted the oestrogen-binding activity from cell extracts. Western blot analysis showed that the antisera specifically recognized the EBP from The antibodies also recognized the protein when the cloned gene was expressed in and did not cross react with proteins. Using electron microscopy and antigen detection by immunogold staining, the EBP appeared to be primarily associated with vacuoles. However, when overexpressed in , the EBP was found diffusely throughout the cell. In conclusion, the EBP has been purified from and antibodies generated against the protein were used to demonstrate that EBP is found associated with vacuoles in

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1995-10-01
2024-03-28
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References

  1. Anaissie E. 1992; Opportunistic mycoses in the immunocompromised host: experience at a cancer center and review.. Clin Infect Dis 14:S43–S53
    [Google Scholar]
  2. Bradford M.M. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.. Anal Biochem 72:248–254
    [Google Scholar]
  3. Crislip M.A., Edwards J.E. 1989; Candidiasis.. Infect Dis Clin N Am (Philadelphia) 3:103–133
    [Google Scholar]
  4. Diamond R.D. 1991; The growing problem of mycoses in patients infected with the human immunodeficiency virus.. Rev Infect Dis 13:480–486
    [Google Scholar]
  5. Frey J., Rohm K.-H. 1978; Subcellular localization and levels of aminopeptidase and dipeptidase in Saccharomyces cerevisiae. . Biochim Biophys Acta 527:31–41
    [Google Scholar]
  6. Kalo A., Witkin S.S. 1989; Candida albicans: cellular immune system interactions during different stages of the menstrual cycle.. Am J Obstet Gynecol 161:1132–1136
    [Google Scholar]
  7. Kinsman O.S., Pitblado K., Coulson C.J. 1988; Effect of mammalian steroid hormones and luteinizing hormone on the germination of Candida albicans and implications for vaginal candidosis.. Mycoses 31:617–626
    [Google Scholar]
  8. Klionsky D.J., Herman P.K., Emr S.D. 1990; The fungal vacuole: composition, function, and biogenesis.. Microbiol Rev 54:266–292
    [Google Scholar]
  9. Laemmli U.K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4.. Nature 227:680–685
    [Google Scholar]
  10. Madani N.D., Malloy P.J., Rodriguez-Pombo P., Krishnan A.V., Feldman D. 1994; Candida albicans estrogen-binding protein gene encodes an oxidoreductase that is inhibited by estradiol.. Proc Natl Acad Sci USA 91922–926
    [Google Scholar]
  11. Mitchell T.G. 1988; Opportunistic mycoses.. In Zinsser Microbiology, 19th edn,. pp. 930–947 Edited by Joklik W. K., Willett H. P., Amos D. B., Wilfert C. M. Norwalk: Appleton & Lange;
    [Google Scholar]
  12. Saito K., Thiele D.J., Davio M., Lockridge O., Massey V. 1991; The cloning and expression of a gene encoding old yellow enzyme from Saccharomyces carlsbergensis. . J Biol Chem 266:20720–20724
    [Google Scholar]
  13. Saral R. 1991; Candida and Aspergillus infections in immunocompromised patients: an overview.. Rev Infect Dis 13:487–492
    [Google Scholar]
  14. Skowronski R., Feldman D. 1989; Characterization of an estrogen-binding protein in the yeast Candida albicans. . Endocrinology 124:1965–1972
    [Google Scholar]
  15. Stott K., Saito K., Thiele D.J., Massey V. 1993; Old yellow enzyme. The discovery of multiple isozymes and a family of related proteins.. J Biol Chem 268:6097–6106
    [Google Scholar]
  16. Yoshihisa T., Anraku Y. 1989; Nucleotide sequence of AMS1, the structure gene of vacuolar α-mannosidase of Saccharomyces cerevisiae. . Biochem Biophys Res Commun 163:908–915
    [Google Scholar]
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