title: Identification and characterization of a zinc metalloprotease associated with invasion by the fish pathogen Vibrio anguillarum.
abstract: An invasiveness-defective mutant of the fish-pathogenic bacterium Vibrio anguillarum was isolated. Compared with the wild type, this mutant had a 1,000-fold higher 50% lethal dose after immersion infection of rainbow trout, Oncorhynchus mykiss, while after intraperitoneal infection, the mutant had only a 10-fold higher 50% lethal dose. In addition, the mutant showed a lower level of protease activity. Two forms of the protease (Pa and Pb) were found after sodium dodecyl sulfate-polyacrylamide gel electrophoresis of nonheated samples. Pa was found predominantly in protease preparations of the wild type, while Pb was the predominant form in the mutant. Conversion of Pb to Pa was observed in protease preparations after incubation at 4 degrees C. Characterization of the protease showed that it was an elastolytic enzyme which required Zn2+ for activity and Ca2+ for stability. The molecular mass of the protease was 36 kilodaltons. N-terminal amino acid sequence analysis of the protease of V. anguillarum revealed homology to the elastase of Pseudomonas aeruginosa and the protease of Legionella pneumophila.
authors.affiliation: Unit of Applied Cell and Molecular Biology, University of Umeå, Sweden.
authors.forename: A
authors.initials: A
authors.lastname: Norqvist
authors.forename: B
authors.initials: B
authors.lastname: Norrman
authors.forename: H
authors.initials: H
authors.lastname: Wolf-Watz
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D004352:Drug Resistance, Microbial; D010450:Endopeptidases / Q000494:pharmacology; D006461:Hemolysis; D008666:Metalloendopeptidases / Q000235:genetics*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009154:Mutation; D014337:Trout / Q000382:microbiology*; D014733:Vibrio / Q000187:drug effects / Q000201:enzymology / Q000235:genetics* / Q000472:pathogenicity; D014774:Virulence; D015032:Zinc / Q000032:analysis*
chemical_list: D010450:Endopeptidases; D008666:Metalloendopeptidases; D015032:Zinc