title: The metalloprotease PrtV from Vibrio cholerae.
abstract: The Vibrio metalloprotease PrtV was purified from the culture supernatant of a Vibrio cholerae derivative that is deficient in several other secreted peptidases, including the otherwise abundant hemagglutinin/protease HapA. The PrtV is synthesized as a 102 kDa protein, but undergoes several N- and C-terminal processing steps during V. cholerae envelope translocation and prolonged incubation. Purified V. cholerae PrtV protease forms of 81 or 73 kDa were stabilized by calcium ions. Removal of calcium resulted in further rapid autoproteolysis. The two major products of autoproteolysis of the PrtV protease were approximately 37 and 18 kDa and could not be separated under non-denaturing conditions, indicating they are interacting domains. In an assay using cultured cells of the human intestinal cell line HCT8, the PrtV protein showed a cytotoxic effect leading to cell death. Using human blood plasma as a source of potential substrates of mammalian origin for the PrtV protease, we found that the extracellular matrix components fibronectin and fibrinogen were degraded by the enzyme. Additional tests with individual protein substrates revealed that plasminogen was also a possible target for the PrtV protease.
authors.affiliation: Department of Molecular Biology, Umeå University, Sweden.
authors.forename: Karolis
authors.initials: K
authors.lastname: Vaitkevicius
authors.forename: Pramod K
authors.initials: PK
authors.lastname: Rompikuntal
authors.forename: Barbro
authors.initials: B
authors.lastname: Lindmark
authors.forename: Rimas
authors.initials: R
authors.lastname: Vaitkevicius
authors.forename: Tianyan
authors.initials: T
authors.lastname: Song
authors.forename: Sun N
authors.initials: SN
authors.lastname: Wai
mesh_terms: D001798:Blood Proteins / Q000378:metabolism; D045744:Cell Line, Tumor; D004795:Enzyme Stability; D006801:Humans; D045726:Metalloproteases / Q000737:chemistry / Q000378:metabolism* / Q000633:toxicity; D010447:Peptide Hydrolases / Q000737:chemistry / Q000378:metabolism* / Q000633:toxicity; D011480:Protease Inhibitors / Q000494:pharmacology; D013379:Substrate Specificity
chemical_list: D001798:Blood Proteins; D011480:Protease Inhibitors; D045726:Metalloproteases; D010447:Peptide Hydrolases; C513132:PrtV protein, Vibrio cholerae