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Characterization of the pili isolated from Vibrio parahaemolyticus O3:K6
N Nakasone1, S Insisengmay, M Iwanaga
1Department of Bacteriology, Faculty of Medicine, University of the Ryukyus, Nishihara, Okinawa, Japan.
Abstract:
Pilus of Vibrio parahaemolyticus O3:K6 strain LVP9 belonging to the newly identified clone was purified and characterized. The molecular mass of the pilin was estimated to be about 18 kDa by SDS-PAGE, and the isoelectric point of the pilin was 5.0 +/- 0.2. The LVP9 pili were antigenically different from the other V. parahaemolyticus Na2 pili and Ha7 pili as previously reported, nevertheless all three had indistinguishable morphology and shared a high degree of homology in their N-terminal amino acid sequences. Strain LVP9 and its purified pili did not agglutinate human and rabbit erythrocytes. The LVP9 organisms and the purified pili were adhesive to the rabbit intestine. The adhesion was inhibited by pretreatment of the rabbit intestine with the purified pili or by pretreatment of the organisms with the Fab fractions of anti-pilus antibody. These results indicate that the LVP9 pilus is an adherent factor to the rabbit intestine.
Insights
The Vibrio parahaemolyticus LVP9 pilus, a key factor in bacterial adhesion, was purified and characterized. This study reveals its role in attaching to the rabbit intestine, highlighting its importance in pathogenesis.
Area of Science:
- Microbiology
- Bacterial Pathogenesis
- Molecular Biology
Background:
- Vibrio parahaemolyticus is a significant foodborne pathogen.
- Pili are known virulence factors in many bacteria.
- Understanding pilus function is crucial for developing control strategies.
Purpose of the Study:
- To purify and characterize the pilus from Vibrio parahaemolyticus O3:K6 strain LVP9.
- To investigate the role of the LVP9 pilus in bacterial adhesion to host tissues.
- To determine the antigenic properties and N-terminal homology of the LVP9 pilus.
Main Methods:
- Pilus purification from Vibrio parahaemolyticus LVP9.
- Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) for molecular mass estimation.
- Isoelectric point determination.
- Antigenic analysis and N-terminal amino acid sequencing.
- In vitro adhesion assays using erythrocytes and rabbit intestinal tissue.
Main Results:
- The pilin molecular mass was approximately 18 kDa with an isoelectric point of 5.0 +/- 0.2.
- LVP9 pili showed distinct antigenic properties but conserved N-terminal homology with other V. parahaemolyticus pili.
- LVP9 pili demonstrated adhesive properties to rabbit intestinal tissue, independent of erythrocyte agglutination.
- Adhesion was specifically inhibited by anti-pilus antibodies, confirming the pilus's role.
Conclusions:
- The LVP9 pilus functions as an adhesin mediating Vibrio parahaemolyticus attachment to the rabbit intestine.
- The pilus is a critical factor in the pathogenesis of Vibrio parahaemolyticus infections.
- Further research into pilus structure-function relationships could yield therapeutic targets.