title: Molecular analysis of an extracellular protease gene from Vibrio parahaemolyticus.
abstract: The structural gene prtVp encoding the extracellular protease of Vibrio parahaemolyticus strain 93 was cloned in Escherichia coli and sequenced. The cloned DNA fragment contained a 1761 bp ORF encoding a 587 amino acid protein. The deduced polypeptide is composed of a 25 amino acid signal peptide and a 562 amino acid extracellular polypeptide with a calculated molecular mass of 63,156 Da. Protease analysis using a gelatin-containing SDS-polyacrylamide gel detected the presence of a 62 kDa protease that was present in the culture supernatant fractions of the wild-type V. parahaemolyticus strain and of E. coli bearing a pUC119 recombinant with the prtVp DNA insert. The protease activity was inhibited by zinc- and metal-specific inhibitors such as EDTA and 1,10-phenanthroline, which suggested that it is a metalloprotease. The deduced amino acid sequence of PrtVp has 32% identity with that of the collagenase of Vibrio alginolyticus, but has no identity with those of the bacterial proteases. A conserved zinc-binding domain was also found in PrtVp from homology comparison with other metalloproteases. This PrtVp can cause weak haemolysis on blood agar.
authors.affiliation: Graduate Institute of Agricultural Chemistry, National Taiwan University, Taipei, Republic of China.
authors.forename: C Y
authors.initials: CY
authors.lastname: Lee
authors.forename: S C
authors.initials: SC
authors.lastname: Su
authors.forename: R B
authors.initials: RB
authors.lastname: Liaw
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics*; D001483:Base Sequence; D001665:Binding Sites; D017124:Conserved Sequence; D005798:Genes, Bacterial; D006461:Hemolysis; D008666:Metalloendopeptidases / Q000096:biosynthesis / Q000187:drug effects / Q000235:genetics*; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D011480:Protease Inhibitors / Q000494:pharmacology; D011994:Recombinant Proteins / Q000096:biosynthesis; D017386:Sequence Homology, Amino Acid; D014733:Vibrio; D014736:Vibrio parahaemolyticus; D015032:Zinc / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D011480:Protease Inhibitors; D011994:Recombinant Proteins; D008666:Metalloendopeptidases; C073584:Prt protein, Streptomyces; D015032:Zinc