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Characterization of outer membrane and secreted proteins of Leptospira interrogans serovar pomona
R L Zuerner1, W Knudtson, C A Bolin
1National Animal Disease Center, Agricultural Research Service, Ames, Iowa 50010.
Abstract:
Outer membrane and secreted proteins were isolated from Leptospira interrogans serovar pomona and characterized by sodium dodecyl sulfate polyacrylamide gel electrophoresis, immunoblot and radioimmunoprecipitation techniques. The L. interrogans outer membranes were extracted with Triton X-114 and contained several proteins. The major cellular protein with a molecular mass of 31 kDa was associated exclusively with the L. interrogans outer membrane. Using a whole cell immunoprecipitation method, five hydrophobic, Triton X-114 extractable proteins (22, 26, 31, 36 and 42 kDa) were exposed on the surface of L. interrogans. The 31 kDa protein was heat labile and was a potent antigen in animals experimentally infected with L. interrogans serovar pomona. Several proteins were secreted by L. interrogans including a 60 kDa protein tentatively identified as the L. interrogans hemolysin.
Insights
Researchers identified key outer membrane and secreted proteins from Leptospira interrogans serovar pomona. A 31 kDa protein was found to be a potent antigen in infected animals, indicating its potential role in leptospirosis.
Area of Science:
- Microbiology
- Immunology
- Protein Chemistry
Background:
- Leptospira interrogans serovar pomona is a significant bacterial pathogen.
- Understanding its surface and secreted proteins is crucial for diagnostics and vaccine development.
Purpose of the Study:
- To isolate and characterize outer membrane and secreted proteins of L. interrogans serovar pomona.
- To identify potential antigens for diagnostic and vaccine applications.
Main Methods:
- Proteins were isolated from L. interrogans outer membranes and secreted fractions.
- Characterization involved sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), immunoblotting, and radioimmunoprecipitation.
- Triton X-114 extraction was used to isolate hydrophobic proteins.
Main Results:
- Several proteins were identified in the outer membrane, including a major 31 kDa protein.
- Five hydrophobic proteins (22, 26, 31, 36, and 42 kDa) were exposed on the bacterial surface.
- The 31 kDa protein was heat-labile and demonstrated potent antigenicity in infected animals.
- A 60 kDa secreted protein, potentially L. interrogans hemolysin, was identified.
Conclusions:
- The study successfully characterized key surface and secreted proteins of L. interrogans serovar pomona.
- The 31 kDa outer membrane protein is a promising candidate antigen for diagnosing and potentially preventing leptospirosis.