This recombinant BrkA autotransporter from Bordetella pertussis is expressed in E. coli and covers the complete sequence from amino acids 1 to 1010. The protein carries dual tags—an N-terminal 10xHis-tag paired with a C-terminal Myc-tag—which makes purification and detection more straightforward. SDS-PAGE analysis confirms purity levels above 85%, suggesting it should perform reliably in most research settings.
BrkA represents an autotransporter protein from Bordetella pertussis that appears central to how this bacterium causes disease. It seems to help create protective barriers that shield the bacteria from host immune attacks, essentially helping it survive longer in infected tissues. Studying BrkA may be important for understanding how bacterial virulence works and could potentially guide development of new treatments for infections caused by Bordetella pertussis.
Potential Applications
Note: The applications listed below are based on what we know about this protein's biological functions, published research, and experience from experts in the field. However, we haven't fully tested all of these applications ourselves yet. We'd recommend running some preliminary tests first to make sure they work for your specific research goals.
Based on the provided information, the recombinant Bordetella pertussis BrkA autotransporter is expressed in E. coli, a prokaryotic system that is generally suitable for producing bacterial proteins like BrkA. As both Bordetella and E. coli are prokaryotes, the protein has a reasonable probability of proper folding. However, BrkA is a complex autotransporter with multiple domains, including passenger and translocation domains that require precise folding and membrane integration. While E. coli can express bacterial membrane proteins, the dual tags (N-terminal 10xHis and C-terminal Myc) may interfere with proper membrane insertion and domain organization. The protein is full-length (1-1010aa) with >85% purity, but since activity is explicitly unverified and autotransporters require specific folding for their secretion and adhesion functions, the protein cannot be assumed to be correctly folded or bioactive without experimental validation.
1. Antibody Development and Immunological Studies
The recombinant BrkA can serve as an effective immunogen for generating antibodies that recognize linear epitopes. The dual tags facilitate purification and detection during antibody screening. However, if BrkA is misfolded, antibodies may not recognize conformational epitopes of native, properly folded BrkA in B. pertussis. Validation against native BrkA from Bordetella cultures is recommended.
2. Protein-Protein Interaction Studies
This application requires caution. While the dual tags enable technical feasibility for pull-down assays, if BrkA is misfolded, it may not interact physiologically with true host cell receptors or bacterial partners. Autotransporter passenger domains require precise conformation for specific adhesion interactions. This application should only be pursued after confirming proper folding through functional assays.
3. Tag-Assisted Purification Method Development
This application is well-suited for technical optimization. The dual tags make this protein useful for developing and refining purification protocols for large bacterial proteins. However, methods developed using this potentially misfolded protein may not directly apply to native, membrane-integrated BrkA.
4. Western Blot and Immunodetection Assay Development
This application is appropriate for detection purposes. The recombinant BrkA can serve as a positive control and standard for immunodetection methods. However, if the protein is misfolded, detection may not correlate with native BrkA in biological samples. Validation against native protein is essential for quantitative applications.
Final Recommendation & Action Plan
Given that BrkA is a bacterial protein expressed in a prokaryotic system, the probability of proper folding is relatively high. However, recommend first performing validation: 1) Biophysical characterization (size-exclusion chromatography, circular dichroism) to assess folding state; 2) Functional validation of autotransporter activity if possible; 3) Comparison with native BrkA from B. pertussis. Antibody development and detection applications can proceed immediately. Interaction studies should await proper folding confirmation. For reliable functional studies, include appropriate controls such as known binding partners and validate findings with native BrkA.
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