Advertisement

cDNA cloning of Candida albicans aspartic proteinase and its diagnostic application

  • S. Y. Murayama
Conference paper
Part of the NATO ASI Series book series (volume 69)

Abstract

The expanding of immunocompromised individuals has focused attention on the importance of Candida species as an opportunistic fungal pathogen. C. albicans is a major infectious fungal agent in humans, invading by attachment to the host cells via cell-surface adhesive molecules. Once attached, C. albicans cells proliferate and release extracellular aspartic proteinase, which aids them to invade the integument of the host. Few extracellular enzymes have been described as virulence factors, but only extracellular an aspartic acid proteinase has received intense attention. There seems to be a relationship between the production of aspartic proteinase by C. albicans and its virulence (7, 9, 13), but the actual role of proteinase as the virulence factor remains unclear. Yamamoto et al. (20) reported that all strains of C. albicans tested secrete proteinase extracellularly, and they sequenced the first 23 amino acids of the N-terminal of the proteinase from their C. albicans No. 114. We have undertaken the cDNA cloning of this enzyme using the polymerase chain reaction (PCR) with mixed primers constructed from the information published by that group. This enzyme has very recently sequenced from the C. albicans strain ATCC10231 (6). We also show the results of comparison of our sequence with theirs and other related enzymes.

Keywords

Candida Albicans Nest Polymerase Chain Reaction Aspartic Proteinase Mixed Primer Aspartyl Proteinase 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Ammerer G, Hunter CP, Rothman JH, Saari GC, Valis LA, Stereng TH (1986) PEP4 gene of Saccharomyces cerevisiae encodes proteinase A, a vacuolar enzyme required for processing of vacuolar precursors. Mol. Cell. Biol. 6:2490–2499.PubMedGoogle Scholar
  2. Andreeva NS, Zdanov AS, Gustchina AE, Federov AA (1984) Structure of ethanol-inhibited porcine pepsin at 2-Å resolution and binding of the methyl ester of phenylalanyl-diiodotyrosine to the enzyme. J. Biol. Chem. 259:11353–11365.PubMedGoogle Scholar
  3. Brown AJP, Bertram G, Feldmann PJF, Peggie MW, Swoboda RK (1991) Codon utilization in the pathogenic yeast, Candida albicans. Nucl. Acids. Res. 19:4298.PubMedCrossRefGoogle Scholar
  4. Faust PL, Kornfelds, S, Chirgwin JM (1985) Cloning and sequence analysis of cDNA for human cathepsin D. Proc. Natl. Acad. Sci. USA 82:4910–4914.PubMedCrossRefGoogle Scholar
  5. Hsu I-N, Delbaere LTJ, James MNG, Hofmann T (1977) Penicillopepsin from Penicillium janthtnellum crystal structure at 2.8 Å and sequence homology with porcine pepsin. Nature 266:140–145.PubMedCrossRefGoogle Scholar
  6. Hube B, Turver CJ, Odds FC, Eiffert H, Boilnois GJ, Küchel H, Rüchel R (1991) Sequence of the Candida albicans gene encoding the secretory aspartate proteinase. J. Med. Vet. Mycol. 29:129–132.PubMedCrossRefGoogle Scholar
  7. Kwon-Chung KJ, Lehman D, Good C, Magee PT (1985) Genetic evidence for role of extracellular proteinase in virulence of Candida albicans. Infect. Immun. 49:571–575.PubMedGoogle Scholar
  8. Lott TJ, Page LS, Boiron P, Bensen J, Reiss E (1989) Nucleotide sequence of the Candida albicans aspartyl proteinase gene. Nucl. Acids Res. 17:1779.PubMedCrossRefGoogle Scholar
  9. MacDonald F, Odds FC. (1983) Virulence for mice of a proteinase-secreting strain of Candida albicans and a proteinase-deficient mutant. J. Gen. Microbiol. 129:431–438.PubMedGoogle Scholar
  10. Mukai H, Takeda O, Asada K, Kato I, Murayama SY, Yamaguchi H (1992) cDNA cloning of an aspartic proteinase secreted by Candida albicans. Microbial Immunol. in press.Google Scholar
  11. Perlman D, Halvorson HO (1983) A putative signal peptidase recognition site and sequence in eukaryotic and prokaryotic signal peptides. J. Mol. Biol. 167:391–409.PubMedCrossRefGoogle Scholar
  12. Rüchel R (1981) Properties of a purified proteinase from the yeast Candida albicans. Biochim. Biophys. Acta 659:99–103.PubMedGoogle Scholar
  13. Sanger F (1981) Determination of nucleotide sequences in DNA. Science 214: 1205–1210.PubMedCrossRefGoogle Scholar
  14. Sripati CE, Warner JR (1978) Isolation, characterization, and translation of mRNA from yeast. In Prescott DM (ed), Methods in Cell Biology, vol 12. Academic Press, New York, p 61–81.CrossRefGoogle Scholar
  15. Subramanian E, Swan IDA, Liu M, Davies DR, Jenkins JA, Tickel IJ, Blundell TL (1977) Homology among acid proteases: comparison of crystal structures at 3 Å resolution of acid proteases from Rhizopus chinensis and Endothia parasitica. Proc. Natl. Acad. Sci. USA 74:556–559.PubMedCrossRefGoogle Scholar
  16. Tang J (1979) Evolution in the structure and function of carboxyl proteases. Mol. Cell. Biochem. 26:93–109.PubMedCrossRefGoogle Scholar
  17. Togni G, Sanglard D, Falchetto R, Monod M (1991) Isolation and nucleotide sequence of the extracellular acid protease gene (ACP) from the yeast Candida tropicalis. FEBS Letters 286:181–185.PubMedCrossRefGoogle Scholar
  18. Takeda O, Mukai H, Takesako K, Sono E, Asada K, Kato I, Murayama SY, Yamaguchi H (1992) Specific detection of Candida albicans strains by amplification of its aspartic proteinase gene using PCR. Microbiol. Immunol, in press.Google Scholar
  19. Yamamoto T, Nohara K, Uchida K, Yamaguchi K (1992) Microbiol. Immunol. 36, in press.Google Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1993

Authors and Affiliations

  • S. Y. Murayama
    • 1
  1. 1.Department of BacteriologyTeikyo University School of MedicineTokyoJapan

Personalised recommendations