1887

Abstract

The nature of secreted aminopeptidases in was investigated by using a reverse genetic approach. genomic and cDNA libraries were screened with spp. and aminopeptidase genes as the probes. Two leucine aminopeptidases, ruLap1 and ruLap2, and two dipeptidyl-peptidases, ruDppIV and ruDppV, were characterized and compared to orthologues secreted by using a recombinant protein from . RuLap1 is a 33 kDa nonglycosylated protein, while ruLap2 is a 58–65 kDa glycoprotein. The hydrolytic activity of ruLap1, ruLap2 and orthologues showed various preferences for different aminoacyl-7-amido-4-methylcoumarin substrates, and various sensitivities to inhibitors and cations. ruDppIV and ruDppV showed similar activities to orthologues. In addition to endopeptidases, the four aminopeptidases ruLap1, ruLap2, ruDppIV and ruDppV were produced by in a medium containing keratin as the sole nitrogen source. Synergism between endo- and exopeptidases is likely to be essential for dermatophyte virulence, since these fungi grow only in keratinized tissues.

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2005-01-01
2024-03-28
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References

  1. Beauvais A., Monod M., Debeaupuis J. P., Diaquin M., Kobayashi H., Latgé J. P. 1997a; Biochemical and antigenic characterization of a new dipeptidyl-peptidase isolated from Aspergillus fumigatus . J Biol Chem 272:6238–6244 [CrossRef]
    [Google Scholar]
  2. Beauvais A., Monod M., Wyniger J., Debeaupuis J. P., Grouzmann E., Brakch N., Svab J., Hovanessian A. G., Latgé J. P. 1997b; Dipeptidyl-peptidase IV secreted by Aspergillus fumigatus , a fungus pathogenic to humans. Infect Immun 65:3042–3047
    [Google Scholar]
  3. Becker J. M., Naider F. 1995; Fungal peptide transport as a drug delivery system. In Peptide-Based Drug Design: Controlling Transport and Metabolism pp 369–384 Edited by Taylor M., Amidon G. Washington, DC: American Chemical Society;
    [Google Scholar]
  4. Beggah S., Foundling S., Monod M, Léchenne B., Reichard U. 2000; Intra and intermolecular events direct the propeptide-mediated maturation of the Candida albicans secreted aspartic proteinase Sap1p. Microbiology 146:2765–2773
    [Google Scholar]
  5. Ben-Meir D., Spungin A., Ashkenazi R., Blumberg S. 1993; Specificity of Streptomyces griseus dinuclear aminopeptidase and modulation of activity by divalent metal ion binding and substitution. Eur J Biochem 212:107–112 [CrossRef]
    [Google Scholar]
  6. Blinkovsky A. M., Byun T., Brown K. M., Golightly E. J., Klotz A. V. 2000; A non-specific aminopeptidase from Aspergillus . Biochim Biophys Acta 1480171–181 [CrossRef]
    [Google Scholar]
  7. Borg-von Zepelin M., Beggah S., Boggian K., Sanglard D., Monod M. 1998; The expression of the secreted aspartyl proteinases Sap4 to Sap6 from Candida albicans in murine macrophages. Mol Microbiol 28:543–554 [CrossRef]
    [Google Scholar]
  8. Brouta F., Descamps F., Fett T., Losson B., Gerday C., Mignon B. 2001; Purification and characterization of a 43·5 kDa keratinolytic metalloprotease from Microsporum canis . Med Mycol 39:269–275 [CrossRef]
    [Google Scholar]
  9. Brouta F., Descamps F., Monod M., Vermout S., Losson B., Mignon B. 2002; Secreted metalloprotease gene family of Microsporum canis . Infect Immun 70:5676–5683 [CrossRef]
    [Google Scholar]
  10. Byun T., Kofod L., Blinkovsky A. 2001; Synergistic action of an X-prolyl dipeptidyl aminopeptidase and a non-specific aminopeptidase in protein hydrolysis. J Food Chem 49:2061–2063 [CrossRef]
    [Google Scholar]
  11. Chambers S. P., Prior S. E., Barstow D. A., Minton N. P. 1988; The pMTL nic cloning vectors. Improved pUC polylinker regions to facilitate the use of sonicated DNA for nucleotide sequencing. Gene 68:139–149 [CrossRef]
    [Google Scholar]
  12. Chien H. C., Lin S. H., Chao S. H., Chen C. C., Wang W. C., Shaw C. Y., Tsai Y. C., Hu H. Y., Hsu W. H. 2002; Purification, characterisation, and genetic analysis of a leucine aminopepeptidase from Aspergillus sojae . Biochim Biophys Acta 1576119–126 [CrossRef]
    [Google Scholar]
  13. Danew P., Friedrich E. 1980; Untersuchung zur Peptidaseaktivität hautpathogener Pilze. IV. Aminopeptidaseaktivität bei Microsporum gypseum und Trichophyton rubrum nach Wachstum in einer Glukose-Salzlösung mit Zusatz von Lysinmonochlorid als N-Quelle. Mycosen 23:502–511
    [Google Scholar]
  14. De Bersaques J., Dockx P. 1973; Proteolytic and leucylnaphthylamidase activity in some dermatophytes. Arch Belg Dermatol Syphiligr 29:135–140
    [Google Scholar]
  15. Descamps F., Brouta F., Monod M., Zaugg C., Baar D., Losson B., Mignon B. 2002; Isolation of a Microsporum canis gene family encoding three subtilisin-like proteases expressed in vivo . J Invest Dermatol 70:830–836
    [Google Scholar]
  16. Doumas A., Affolter M., Monod M, Van den Broek P. 1998; Characterization of the prolyl dipeptidyl peptidase gene ( dppIV ) from the koji mold Aspergillus oryzae . Appl Environ Microbiol 64:4809–4815
    [Google Scholar]
  17. Ellis K. J., Morrison J. F. 1982; Buffers of constant ionic strength for studying pH-dependent processes. Methods Enzymol 87:405–426
    [Google Scholar]
  18. Greenblatt H. M., Almog O., Maras B., Spungin-Bialik A., Barra D., Blumberg S., Shoham G. 1997; Streptomyces griseus dinuclear aminopeptidase: X-ray crystallographic structure at 1·75 Å resolution. J Mol Biol 212:620–636
    [Google Scholar]
  19. Grossberger D. 1987; Minipreps of DNA from bacteriophage lambda. Nucleic Acids Res 15:6737 [CrossRef]
    [Google Scholar]
  20. Hall T. A. 1999; BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symp Ser 41:95–98
    [Google Scholar]
  21. Hasselgren C., Park H. I., Ming L. J. 2001; Metal ion binding and activation of Streptomyces griseus dinuclear aminopeptidase: cadmium(II) binding as a model. J Biol Inorg Chem 6:120–127 [CrossRef]
    [Google Scholar]
  22. Hauser M., Narita V., Donhardt A. M., Naider F., Becker J. M. 2001; Multiplicity and regulation of genes encoding peptide transporters in Saccharomyces cerevisiae . Mol Membr Biol 18:105–112 [CrossRef]
    [Google Scholar]
  23. Jousson O., Capoccia S., Mignon B., Barblan J., Quadroni M., Monod M, Léchenne B., Bontems O. 2004a; Multiplication of an ancestral gene encoding secreted fungalysin preceded species differentiation in the dermatophytes Trichophyton and Microsporum . Microbiology 150:301–310 [CrossRef]
    [Google Scholar]
  24. Jousson O., Mignon B., Reichard U., Barblan J., Quadroni M., Monod M, Léchenne B., Bontems O. 2004b; Secreted subtilisin gene family in Trichophyton rubrum . Gene 339:79–88 [CrossRef]
    [Google Scholar]
  25. Julius D., Brake A., Blair L., Kunisawa R., Thorner J. 1984; Isolation of the putative structural gene for the lysine-arginine-cleaving endopeptidase required for processing of yeast prepro- α -factor. Cell 37:1075–1089 [CrossRef]
    [Google Scholar]
  26. Kwong-Chung K. J., Bennet J. E. 1992 Medical Mycology Philadelphia & London: Lea & Febiger;
    [Google Scholar]
  27. Lin L.-Y., Park H. I., Ming L. J. 1997; Metal-binding and active-site structure of di-zinc Streptomyces griseus aminopeptidase. J Biol Inorg Chem 2:744–749 [CrossRef]
    [Google Scholar]
  28. Lubkowitz M. A., Hauser L., Breslav M., Naider F., Becker J. M. 1997; An oligopeptide transport gene from Candida albicans . Microbiology 143:387–396 [CrossRef]
    [Google Scholar]
  29. Mignon B., Swinnen M., Bouchara J., Hofinger M., Nikkels A., Pierard G., Gerday C., Losson B. 1998; Purification and characterization of a 31·5 kDa keratinolytic subtilisin-like serine protease from Microsporum canis and evidence of its secretion in naturally infected cats. Med Mycol 36:395–404 [CrossRef]
    [Google Scholar]
  30. Monod M., Applegate L. A. 1994; RNA extractions and Northern hybridisation of Aspergillus fumigatus for the detection of alkaline and metalloprotease gene expression. In Molecular Biology of Pathogenic Fungi: a Laboratory Manual pp 29–32 Edited by Maresca B., Kobayashi G. S. New York: Telos Press;
    [Google Scholar]
  31. Monod M., Togni G., Rahalison L., Frenk E. 1991; Isolation and characterisation of an extracellular alkaline protease of Aspergillus fumigatus . J Med Microbiol 35:23–28 [CrossRef]
    [Google Scholar]
  32. Monod M., Togni G., Hube B., Sanglard D. 1994; Multiplicity of genes encoding secreted aspartic proteinases in Candida species. Mol Microbiol 13:357–368 [CrossRef]
    [Google Scholar]
  33. Monod M., Jaton-Ogay K., Reichard U. 1999; Aspergillus fumigatus secreted proteases as antigenic molecules and virulence factors. Contrib Microbiol 2:182–192
    [Google Scholar]
  34. Monod M., Jaccoud S., Zaugg C., Panizzon R, Léchenne B., Baudraz F. 2002; Survey of dermatophyte infections in Lausanne area (Switzerland). Dermatology 205:201–203 [CrossRef]
    [Google Scholar]
  35. Nakadai T., Nasuno S., Iguchi N. 1973; Purification and properties of leucine aminopeptidase I from Aspergillus oryzae . Agric Biol Chem 37:757–765 [CrossRef]
    [Google Scholar]
  36. Nishizawa M., Yasuhara T., Nakai T., Fujiki Y., Ohashi A. 1994; Molecular cloning of the aminopeptidase Y gene of Saccharomyces cerevisiae . J Biol Chem 269:13651–13655
    [Google Scholar]
  37. O'Cuinn G., Fitzgerald R., Bouchier P., McDonnell M. 1999; Generation of non-bitter casein hydrolysates by using combinations of a proteinase and aminopeptidases. Biochem Soc Trans 27:730–734
    [Google Scholar]
  38. Payne J. W., Smith M. W. 1994; Peptide transport by microorganisms. Adv Microbiol Physiol 36:1–80
    [Google Scholar]
  39. Rubio-Aliaga I., Daniel H. 2002; Mammalian peptide transporters as targets for drug delivery. Trends Pharmacol Sci 23:434–440 [CrossRef]
    [Google Scholar]
  40. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual , 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  41. Stacey G., Koh S., Granger C., Becker J. M. 2002; Peptide transport in plants. Trends Plant Sci 80:270–279
    [Google Scholar]
  42. Toma C., Honma Y. 1996; Cloning and genetic analysis of the Vibrio cholerae aminopeptidase gene. Infect Immun 64:4495–4500
    [Google Scholar]
  43. Weitzman I., Summerbell R. C. 1995; The dermatophytes. Clin Microbiol Rev 8:240–259
    [Google Scholar]
  44. Woodfolk J. A., Wheatley L. M., Piyasena R. V., Benjamin D. C., Platts-Mills T. A. 1998; Trichophyton antigens associated with IgE antibodies and delayed-type hypersensitivity - sequence homology to two families of serine proteinases. J Biol Chem 273:29489–29496 [CrossRef]
    [Google Scholar]
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