Borrelia Spielmanii DbpA
Description
Recombinant Borrelia Spielmanii Decorin Binding Protein A produced in E.coli is a non-glycosylated, polypeptide chain having a calculated molecular mass of 17kDa.
Borrelia Spielmanii DbpA is expressed with a -10x His tag at N-terminus and purified by proprietary chromatographic techniques.
Product Specs
Borrelia, a bacterial genus within the spirochete phylum, is responsible for causing borreliosis. This zoonotic vector-borne disease is primarily transmitted by ticks, with some species also transmitted by lice. Among the 36 identified Borrelia species, 12 are known to cause Lyme disease or borreliosis and are spread through tick bites. Borreliella spielmanii, a Gram-negative bacterium, is one of the pathogens belonging to the Borreliella burgdorferi sensu lato complex, which causes Lyme disease. DbpA, also referred to as p17 or Osp17, exhibits heterogeneity among the human pathogenic B. burgdorferi sensu lato species.
Recombinant Borrelia Spielmanii Decorin Binding Protein A, produced in E.coli, is a non-glycosylated polypeptide chain with a calculated molecular mass of 17kDa.
A -10x His tag is added to the N-terminus of Borrelia Spielmanii DbpA during expression, and purification is achieved using proprietary chromatographic techniques.
Borrelia Spielmanii DbpA is supplied in a buffer solution containing 20mM HEPES (pH 7.6), 250mM NaCl, and 20% glycerol.
1. Exhibits binding affinity for human IgG and IgM antibodies.
2. Applicable for use in immunodot testing with plasma samples from individuals with positive or negative Lyme disease status.
Q&A
What is Borrelia spielmanii and how is it related to Lyme disease?
Borrelia spielmanii (formerly designated genospecies A14S) is a human pathogenic genospecies within the Borrelia burgdorferi sensu lato complex that causes Lyme disease in Europe . This spirochete has been isolated from patients with skin manifestations in The Netherlands, Germany, Denmark, Hungary, and Slovenia . B. spielmanii belongs to the genus of bacteria of the spirochete phylum and is primarily transmitted by ticks . It is one of at least 12 Borrelia species distinguished to cause Lyme disease out of the 36 known species of Borrelia .
What is DbpA and what is its significance in Borrelia spielmanii research?
DbpA (Decorin Binding Protein A), also known as p17 or Osp17, is a heterogeneous protein found among the human pathogenic B. burgdorferi sensu lato species, including B. spielmanii . This protein has a calculated molecular mass of 17kDa and plays important roles in bacterial adhesion to host tissues through its binding to decorin, a proteoglycan associated with collagen fibers. DbpA also demonstrates immunological functions, as it binds to IgG- and IgM-type human antibodies , suggesting its involvement in host-pathogen interactions during infection.
Where is the dbpA gene located in the B. spielmanii genome?
The dbpA gene in B. spielmanii is located on linear plasmids. Analysis of four human B. spielmanii isolates revealed variation in plasmid profiles, with the dbpA gene located on plasmids of different sizes . Three isolates carried the dbpA gene on a 50 kb linear plasmid, while one isolate (PSigII) contained the gene on a larger 100 kb plasmid . This variation may be related to host-pathogen interactions, as the PSigII strain with the larger plasmid was isolated from a patient with a previous history of Lyme disease .
What techniques are recommended for isolation and purification of B. spielmanii DbpA?
Based on the research literature, recombinant B. spielmanii DbpA can be effectively produced in E. coli expression systems . The protein is typically expressed with a 10x His tag at the N-terminus to facilitate purification using chromatographic techniques . The recombinant protein is non-glycosylated and has a calculated molecular mass of 17kDa . For optimal stability, the purified protein should be maintained in 20mM HEPES buffer (pH-7.6) containing 250mM NaCl and 20% glycerol . Multiple freeze-thaw cycles should be avoided to maintain protein stability and function .
How can researchers assess DbpA expression in different B. spielmanii isolates?
To assess DbpA expression across different B. spielmanii isolates, researchers should consider a multi-method approach:
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Pulsed-field gel electrophoresis (PFGE): This technique has been successfully used to analyze B. spielmanii isolates and localize the dbpA gene on linear plasmids . PFGE allows visualization of large DNA molecules and can reveal differences in plasmid profiles between isolates.
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Western blotting: This technique can detect DbpA protein expression in different isolates, as demonstrated with other Borrelia proteins . Protein extracts from different isolates can be separated by SDS-PAGE and probed with specific antibodies against DbpA.
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Immunofluorescence microscopy: This method could be adapted to visualize DbpA expression on the surface of B. spielmanii cells, similar to approaches used for detecting complement components on Borrelia .
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qRT-PCR: While not specifically mentioned in the search results, quantitative reverse transcription PCR could provide information about dbpA gene expression levels across different isolates.
These complementary approaches would provide comprehensive data on both gene presence and protein expression levels.
How does B. spielmanii DbpA interact with the human immune system?
B. spielmanii DbpA exhibits specific interactions with components of the human immune system. Most notably, it binds to IgG- and IgM-type human antibodies , indicating its recognition by the adaptive immune response during infection. This binding can be demonstrated through immunodot tests with Lyme disease positive/negative plasma samples .
While the direct role of DbpA in complement evasion is not fully characterized in the available research, B. spielmanii as a species has developed mechanisms to resist complement-mediated killing . The bacterium acquires host alternative pathway regulators factor H and factor H-like protein (FHL-1) from human serum, which retain their factor I-mediated C3b inactivation activities when bound to the spirochetal surface . This acquisition is mediated by surface-exposed proteins designated BsCRASP-1 to -3 . Future research may reveal potential interactions between DbpA and these complement evasion processes.
Why do different B. spielmanii isolates show variation in plasmid profiles carrying the dbpA gene?
The variation in plasmid profiles among B. spielmanii isolates represents an interesting area of research with potential implications for pathogenesis and host adaptation. Analysis of four human B. spielmanii isolates revealed that one isolate (PSigII) contained a 100 kb plasmid carrying the dbpA gene, while three other isolates had the gene on a 50 kb plasmid .
This variation may be a result of host-pathogen interactions and bacterial adaptation. Notably, the PSigII strain with the larger plasmid was isolated from a patient with a previous history of Lyme disease, whereas the other three isolates came from patients diagnosed with Lyme disease for the first time . This correlation suggests that plasmid variations might be associated with differences in pathogenicity, persistence, or immune evasion capabilities.
Potential explanations for these variations include:
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Genomic rearrangements in response to immune pressure
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Acquisition of additional genetic material through horizontal gene transfer
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Adaptations that enhance survival in specific host environments
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Selection for variants with altered expression of surface proteins like DbpA
How can B. spielmanii DbpA protein be used in serological tests for Lyme disease?
Given that B. spielmanii DbpA binds to IgG- and IgM-type human antibodies , this protein has potential applications in serological diagnostics for Lyme disease. Researchers can exploit this property to develop or improve diagnostic tests in the following ways:
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Immunodot tests: B. spielmanii DbpA can be used in immunodot assays with patient plasma to detect Borrelia-specific antibodies . This approach could potentially distinguish between Lyme disease positive and negative samples.
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ELISA-based diagnostics: While not directly mentioned in the search results, recombinant DbpA could be employed as an antigen in enzyme-linked immunosorbent assays for detecting patient antibodies against Borrelia.
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Multiplex serological arrays: DbpA could be included in panels of Borrelia antigens to improve the sensitivity and specificity of Lyme disease diagnostics, particularly for infections caused by B. spielmanii.
When developing such tests, researchers should consider the potential heterogeneity of DbpA across different B. spielmanii isolates and optimize conditions for recombinant protein stability.
What is the relationship between DbpA and complement resistance mechanisms in B. spielmanii?
B. spielmanii demonstrates variable resistance to complement-mediated killing, with most isolates showing high levels of serum resistance . While the direct role of DbpA in complement evasion remains to be fully characterized, the bacterium employs several mechanisms to resist complement:
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Acquisition of host complement regulators: B. spielmanii isolates acquire the host alternative pathway regulators factor H and FHL-1 from human serum . These regulators maintain their factor I-mediated C3b inactivation activities when bound to the bacterial surface.
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Expression of complement regulator-acquiring surface proteins: B. spielmanii expresses proteins designated BsCRASP-1 to -3 (approximately 23-25 kDa in mass) that bind factor H and FHL-1 . Different isolates express distinct patterns of these proteins, with tick isolate PC-Eq17 uniquely expressing BsCRASP-3 .
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Differential complement activation: All B. spielmanii isolates activate the complement system (as shown by C3 deposition), but resistant isolates limit the deposition of later activation products C6 and C5b-9 (terminal complement complex) .
Future research could investigate potential interactions between DbpA and these complement evasion mechanisms, particularly whether DbpA influences the binding of complement regulators or the assembly of the terminal complement complex.
What are the structural and biochemical properties of recombinant B. spielmanii DbpA?
The available research provides several key properties of recombinant B. spielmanii DbpA:
These properties provide important considerations for researchers working with recombinant B. spielmanii DbpA in experimental settings.
How does the serum resistance of B. spielmanii isolates correlate with their expression of surface proteins?
Research has revealed interesting correlations between serum resistance and surface protein expression in B. spielmanii isolates:
This data shows that serum resistance in B. spielmanii correlates with the expression of specific complement regulator-acquiring surface proteins (CRASPs) and the ability to limit deposition of terminal complement components. Further research is needed to determine if DbpA interacts with these mechanisms or contributes independently to immune evasion.
Product Science Overview
Borrelia spielmanii is a species of spirochete bacteria belonging to the Borrelia burgdorferi sensu lato complex, which is known to cause Lyme borreliosis. This species was identified relatively recently and is one of the several Borrelia species that can infect humans. Decorin binding protein A (DbpA) is a surface adhesin found in Borrelia species, including Borrelia spielmanii. DbpA plays a crucial role in the pathogenesis of Lyme disease by facilitating the attachment of the bacteria to host tissues. The recombinant form of this protein, Borrelia spielmanii Decorin Binding Protein A Recombinant, is produced using genetic engineering techniques to study its structure, function, and potential as a target for vaccines and diagnostics.
The recombinant Borrelia spielmanii Decorin Binding Protein A is typically produced in a bacterial expression system, such as Escherichia coli (E. coli). The gene encoding DbpA is cloned into an expression vector, which is then introduced into E. coli cells. These cells are cultured under conditions that induce the expression of the recombinant protein. The protein is often tagged with a histidine (His) tag to facilitate purification. After expression, the cells are lysed, and the recombinant protein is purified using affinity chromatography techniques, such as nickel-nitrilotriacetic acid (Ni-NTA) chromatography, which binds to the His tag. The purified protein is then analyzed for purity and functionality .
Decorin binding protein A (DbpA) interacts with decorin, a collagen-associated proteoglycan found in the extracellular matrix of host tissues. This interaction is critical for the adhesion of Borrelia spielmanii to host cells, which is a key step in the establishment of infection. The binding of DbpA to decorin can be studied using various biochemical and biophysical techniques, such as surface plasmon resonance (SPR) and enzyme-linked immunosorbent assays (ELISA). These methods allow researchers to quantify the binding affinity and kinetics of the interaction between DbpA and decorin.
Additionally, the immunogenicity of DbpA can be analyzed by examining the antibody response in infected individuals or animals. Studies have shown that DbpA is highly immunogenic and elicits a strong antibody response, making it a potential candidate for vaccine development . The recombinant form of DbpA can be used to generate specific antibodies, which can then be employed in diagnostic assays to detect Borrelia spielmanii infections.
