Inhibition of the keratinolytic subtilisin protease Sub3 from Microsporum canis by its propeptide (proSub3) and evaluation of the capacity of proSub3 to inhibit fungal adherence to feline epidermis.
Microsporum canis is a pathogenic fungus that causes a superficial cutaneous infection called dermatophytosis, mainly in cats, dogs and humans. Proteolytic enzymes have been postulated to be key factors involved in the invasion of the stratum corneum and keratinized epidermal structures. Among these proteases, the secreted subtilisin protease Sub3 was found to be required for adherence of M. canis arthroconidia to feline epidermis. This protease is synthetized as a preproenzyme consisting of a signal peptide followed by the propeptide and the protease domain. In order to assess whether the enzymatic activity of Sub3 could be responsible for the role of the protease in the adherence process, we expressed and characterized the propeptide of Sub3 and demonstrated that this propeptide is a strong inhibitor of its mature enzyme. This propeptide acts as a noncompetitive inhibitor with dissociation constants, K(I) and [Formula: see text] of 170 and 130 nM respectively. When tested for its capacity to inhibit adherence of M. canis to feline epidermis using an ex vivo adherence model made of feline epidermis, the propeptide does not prevent adherence of M. canis arthroconidia because it loses its capacity to inhibit rSub3 following a direct contact with living arthroconidia, presumably through inactivation by fungal membrane-bound proteases.
Department of Parasitology and Parasitic Diseases, Faculty of Veterinary Medicine, University of Liège, B43 Sart Tilman, 4000 Liège, Belgium.
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A
Baldo
A
A
Chevigné
M-E
ME
Dumez
A
A
Mathy
P
P
Power
J
J
Tabart
L
L
Cambier
M
M
Galleni
B
B
Mignon
D000818:Animals; D002371:Cat Diseases / Q000382:microbiology*; D002415:Cats; D003881:Dermatomycoses / Q000382:microbiology / Q000473:pathology / Q000662:veterinary*; D004792:Enzyme Precursors / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D004817:Epidermis / Q000382:microbiology* / Q000473:pathology; D004926:Escherichia coli / Q000235:genetics; D066298:In Vitro Techniques; D008864:Microsporum / Q000201:enzymology / Q000472:pathogenicity / Q000502:physiology*; D010447:Peptide Hydrolases / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000494:pharmacology; D020860:Subtilisin / Q000737:chemistry
D004792:Enzyme Precursors; D011994:Recombinant Proteins; D010447:Peptide Hydrolases; D020860:Subtilisin