Recombinant expression and antigenic properties of a 31.5-kDa keratinolytic subtilisin-like serine protease from Microsporum canis.
A secreted 31.5-kDa keratinolytic subtilase (SUB3; AJ431180) is thought to be a Microsporum canis virulence factor and represents a candidate for vaccination trials. In this study, the recombinant keratinase (r-SUB3) was produced by the Pichia pastoris expression system and purified to homogeneity. Recombinant SUB3 displayed identical biochemical properties with the native protease. Experimentally cutaneously infected guinea pigs showed specific lymphoproliferative response towards r-SUB3, while no specific humoral immune response was induced except for one animal. The heterologous expression of SUB3 provides a valuable tool for addressing further investigations on the role of this keratinase in the specific cellular immune response and on its use in vaccination trials in the cat.
Descamps
Frédéric
F
Department of Parasitology and Parasitic Diseases, Faculty of Veterinary Medicine, University of Liège, B-43 Sart-Tilman, 4000 Liège, Belgium.
Brouta
Frédéric
F
Vermout
Sandy
S
Monod
Michel
M
Losson
Bertrand
B
Mignon
Bernard
B
D000818:Animals; D000908:Antibodies, Fungal / Q000097:blood; D000946:Antigens, Fungal / Q000276:immunology*; D015870:Gene Expression; D006168:Guinea Pigs; D007633:Keratins / Q000378:metabolism; D008864:Microsporum / Q000201:enzymology* / Q000235:genetics; D010843:Pichia / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis; D012697:Serine Endopeptidases / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D020860:Subtilisin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013381:Subtilisins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*
D000908:Antibodies, Fungal; D000946:Antigens, Fungal; D011994:Recombinant Proteins; D007633:Keratins; D012697:Serine Endopeptidases; D013381:Subtilisins; C114518:keratinolytic subtilase; D020860:Subtilisin