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Microbiology 151 (2005), 145-155; DOI  10.1099/mic.0.27484-0
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Microbiology 151 (2005), 145-155; DOI  10.1099/mic.0.27484-0
© 2005 Society for General Microbiology

Aminopeptidases and dipeptidyl-peptidases secreted by the dermatophyte Trichophyton rubrum

Michel Monod1, Barbara Léchenne1, Olivier Jousson1, Daniela Grand2, Christophe Zaugg1, Reto Stöcklin3 and Eric Grouzmann2

1 Service de Dermatologie et Vénéréologie, Centre Hospitalier Universitaire Vaudois, Lausanne, Switzerland
2 Division de Pharmacologie et Toxicologies Cliniques, Centre Hospitalier Universitaire Vaudois, Lausanne, Switzerland
3 Atheris Laboratories, case postale 314, Bernex-Genève, Switzerland

Correspondence
Michel Monod
Michel.Monod{at}chuv.hospvd.ch

The nature of secreted aminopeptidases in Trichophyton rubrum was investigated by using a reverse genetic approach. T. rubrum genomic and cDNA libraries were screened with Aspergillus spp. and Saccharomyces cerevisiae 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 Aspergillus fumigatus using a recombinant protein from Pichia pastoris. RuLap1 is a 33 kDa nonglycosylated protein, while ruLap2 is a 58–65 kDa glycoprotein. The hydrolytic activity of ruLap1, ruLap2 and A. fumigatus 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 A. fumigatus orthologues. In addition to endopeptidases, the four aminopeptidases ruLap1, ruLap2, ruDppIV and ruDppV were produced by T. rubrum 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.

Abbreviations: AMC, 7-amido-4-methylcoumarin; E-64, L-trans-epoxysuccinyl-Leu-4-guanidinobutylamide; TLCK, N-p-tosyl-L-lysine chloromethyl ketone; TPCK, N-tosyl-L-phenylalanine chloromethyl ketone

The GenBank/EMBL/DDBJ accession numbers for the nucleotide sequences reported in this paper are AY496930, AY436356, AY496929, AY436357, AY497021, U87950, AF407232 and L48074.




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