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J. Bacteriol. doi:10.1128/JB.01039-07
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Identification of a Vibrio furnissii Oligopeptide Permease and Characterization of Its In Vitro Hemolytic Activity

Department of Biological Science and Technology, National Chiao Tung University, Hsin-Chu, Taiwan, 300, Republic of China

^* To whom correspondence should be addressed. Email: tkwmll{at}mail.nctu.edu.tw.

	Abstract

We describe the purification and characterization of an oligopeptide permease protein (Hly-OppA), from Vibrio furnissii that possesses multi-faceted functions in solute-binding, in vitro hemolysis, antibiotic resistance, and as a virulence factor in bacterial pathogenesis. The solute-binding identity was revealed by N-terminal and internal peptide sequences of the purified protein and confirmed by discernible effects on oligopeptide binding, accumulation of fluorescent substrate, and fluorescent substrate/antibiotic competition assay experiments. The purified protein exhibited host-specific in vitro hemolytic activity against various mammalian erythrocytes and apparent cytotoxicity in CHO-K1 cells. The recombinant Hly-OppA protein and an anti-Hly-OppA monoclonal antibody also exhibited and neutralized the in vitro hemolytic activity, respectively, which further confirms the hemolytic activity of the gene product. In addition, a V. furnissii hly-oppA knockout mutant caused less mortality than the wild-type strain when inoculated into BALB/c mice, indicating the virulence function of this protein. Finally, the in vitro hemolytic activity was also confirmed in homologous ABC-type transporter proteins from other Vibrio species.