[Characterization of an extracellular protease from Clostridium difficile].
Clostridium difficile is an intestinal pathogen, which produces two main virulence factors, the exotoxins A and B. Other bacterial structures have been implicated in the colonization of the gastrointestinal tract, which is the first step of the pathogenic process. C. difficile expresses adherence factors and also, displays some surface-associated proteolytic activity, which could play a role in the physiopathology of this bacterium. The aim of this work was to study the protein named Cwp84 which displays significant homologies with many cysteine proteases. The coding catalytic domain of this protein has been cloned in the expression system pGEX-6P-1, as an in-frame fusion with the gluthatione S-transferase, and subsequently purified. The purified fraction showed proteolytic activity on gelatine and BAPNA, but not on azocoll, suggesting a highly selective substrate specificity. The results obtained from inhibition experiments confirmed that Cwp84 belongs to the cysteine protease family. Cwp84 could play a role in degrading some specific host proteins or in the maturation of surface-associated bacterial proteins.
Janoir
C
C
Département de microbiologie-EA 3534, faculté de pharmacie, université de Paris Sud, 5, rue Jean-Baptiste-Clément, 92296 Châtenay-Malabry cedex, France. claire.janoir@cep.u-psud.fr
Grénery
J
J
Savariau-Lacomme
M-P
MP
Collignon
A
A
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D016360:Clostridioides difficile / Q000201:enzymology* / Q000472:pathogenicity; D017124:Conserved Sequence; D003546:Cysteine Endopeptidases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology; D005110:Extracellular Space / Q000201:enzymology; D008969:Molecular Sequence Data; D010447:Peptide Hydrolases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D014774:Virulence
D010447:Peptide Hydrolases; D003546:Cysteine Endopeptidases