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Immuno-fluorescence Assay of Leptospiral Surface-exposed Proteins
Published on: July 1, 2011
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A protein-protein interaction dictates Borrelial infectivity.
Meghna Thakur1, Kavita Sharma1, Kinlin Chao2
1Department of Veterinary Medicine, University of Maryland, College Park and Virginia-Maryland Regional College of Veterinary Medicine, College Park, MD, USA.
Scientific Reports
|June 9, 2017
Summary
The interaction between Borrelia burgdorferi proteins BB0238 and BB0323 is crucial for Lyme disease transmission. Disrupting this interaction prevents spirochete infection in mammals, offering a potential target for new therapies.
Area of Science:
- Microbiology
- Molecular Biology
- Infectious Diseases
Background:
- Borrelia burgdorferi, the causative agent of Lyme disease, possesses complex protein interactions influencing its survival and infectivity.
- The specific roles and significance of interactions between B. burgdorferi proteins BB0238 and BB0323 have remained largely unknown.
Purpose of the Study:
- To identify the specific protein segments involved in the interaction between BB0238 and BB0323.
- To investigate the functional impact of this interaction on B. burgdorferi infectivity and transmission.
Main Methods:
- Polypeptide segment identification for BB0238-BB0323 binding.
- Analysis of protein structure, stability, and oligomerization.
- Generation and assessment of mutant B. burgdorferi isolates lacking specific interaction domains.
- Luciferase assay for studying interaction inhibition.
Main Results:
- The interaction involves amino acid residues 22-200 in BB0323 and a crucial 11-amino acid segment (120-130) in BB0238.
- Deletion of the BB0238 interaction region destabilizes BB0323 post-translationally.
- Mutant B. burgdorferi lacking this interaction persist in ticks but fail to infect mice, indicating impaired transmission and infectivity.
- A luciferase assay was established to monitor BB0238-BB0323 interaction.
Conclusions:
- The BB0238-BB0323 interaction is essential for the post-translational stability of BB0323.
- This interaction is critical for the mammalian infectivity and transmission of Borrelia burgdorferi.
- The developed luciferase assay provides a platform for discovering small molecule inhibitors against B. burgdorferi infection.
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