Recombinant Bartonella henselae Probable periplasmic serine endoprotease DegP-like (htrA) is expressed in E. coli, spanning amino acids 19-503 of the mature protein. This product carries an N-terminal 10xHis-tag and a C-terminal Myc-tag for streamlined purification and detection. The protein achieves greater than 85% purity, as confirmed by SDS-PAGE, which appears to provide high-quality material for research applications.
The DegP-like serine endoprotease from Bartonella henselae likely plays a crucial role in protein quality control within the periplasmic space. It's involved in degrading misfolded proteins and maintaining cellular homeostasis, functioning as part of the serine protease family. This protein has drawn research interest particularly in studies of bacterial proteolytic processes and stress response mechanisms.
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 Bartonella henselae HtrA protein is expressed in E. coli, a prokaryotic system that is generally suitable for producing bacterial proteins like HtrA. As a bacterial protein expressed in its native prokaryotic environment, the probability of proper folding is relatively high. HtrA is a serine protease/chaperone that requires precise folding for its proteolytic and chaperone functions. The protein is expressed as the mature form (19-503aa) with dual tags (N-terminal 10xHis and C-terminal Myc) and >85% purity. However, HtrA proteins typically form complex oligomeric structures (usually hexamers) and require proper active site formation for protease activity. Since activity is unverified, the protein cannot be assumed to be correctly folded or bioactive without experimental validation of its proteolytic activity and oligomeric state.
1. Antibody Development and Immunological Studies
The recombinant HtrA can serve as an effective immunogen for generating antibodies that recognize linear epitopes. The dual tags facilitate purification and detection. However, antibodies may not recognize conformational epitopes of properly oligomerized native HtrA. Validation against native HtrA from B. henselae is recommended.
2. Protein-Protein Interaction Studies
This application requires caution. While the tags enable technical feasibility for pull-down assays, if HtrA is misfolded or improperly oligomerized, it may not interact physiologically with true substrates or binding partners. HtrA requires proper oligomerization for its chaperone functions and substrate recognition. This application should only be pursued after confirming proper folding and oligomeric state.
3. Comparative Biochemical Analysis
This application is well-suited but requires validation. Basic biochemical characterization is feasible, but comparative studies with HtrA homologs require proper folding to yield valid evolutionary and functional insights. The protein's oligomeric state and stability should be confirmed before meaningful comparisons can be made.
4. ELISA-Based Detection Assays
This application is appropriate for detection purposes. The dual tags enable technical development of ELISA formats for antibody detection. However, if the HtrA protein is misfolded, conformational epitopes may not be properly presented, potentially affecting detection sensitivity for antibodies targeting discontinuous epitopes.
Final Recommendation & Action Plan
Given that this is a bacterial protein expressed in a prokaryotic system, the probability of proper folding is relatively high. However, we recommend first validating the HtrA protein's proteolytic activity using known HtrA substrates and confirming its oligomeric state through size-exclusion chromatography with multi-angle light scattering. Antibody development can proceed as the safest application. For interaction studies and comparative analyses, await validation of proper folding and oligomerization. Always include appropriate controls such as protease inhibitors and known substrates in activity assays. If proper folding and activity are confirmed, the protein becomes suitable for all described applications.
