Recombinant Staphylococcus aureus Gamma-hemolysin component A (hlgA) is produced in E. coli and includes the complete mature protein sequence, spanning amino acids 30 to 309. The design incorporates an N-terminal 10xHis-tag alongside a C-terminal Myc-tag, which streamlines both purification and detection processes. SDS-PAGE analysis confirms protein purity exceeds 90%, making it appropriate for diverse research applications.
Gamma-hemolysin component A belongs to the bi-component leukotoxin family that Staphylococcus aureus produces. The protein appears to play a significant role in creating pores within host cell membranes, which likely contributes to the bacterium's pathogenic properties. Working together with other toxin components, it may disrupt normal cellular functions—offering researchers important clues about how bacteria cause disease and interact with their hosts.
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 Staphylococcus aureus Gamma-hemolysin component A (hlgA) is expressed in E. coli, a prokaryotic system that is generally suitable for producing bacterial proteins like hlgA. As a bacterial toxin, hlgA has a high probability of correct folding in E. coli, which shares a similar prokaryotic environment. The protein is full-length mature (30-309aa) with dual tags (N-terminal 10xHis and C-terminal Myc) and high purity (>90%). However, since activity is explicitly unverified and the tags may potentially interfere with proper folding or oligomerization (as hlgA may require complex formation with other components for hemolytic activity), the protein cannot be assumed to be correctly folded or bioactive without experimental validation of its hemolytic function or binding capabilities.
1. Antibody Development and Immunoassay Research
The recombinant hlgA can serve as an effective immunogen for generating antibodies that recognize linear epitopes, even if the protein is misfolded. The dual tags facilitate purification and detection during screening. However, if hlgA is misfolded, antibodies may not recognize conformational epitopes of native, properly folded hlgA in S. aureus. Validation against native hlgA from bacterial cultures is recommended to ensure physiological relevance.
2. Protein-Protein Interaction Studies
This application requires caution. While the dual tags enable technical feasibility for pull-down assays, if hlgA is misfolded, it may not interact physiologically with true binding partners (e.g., other hemolysin components or host cell receptors). Identified interactions could be non-physiological artifacts due to tag interference or misfolding. This application should only be pursued after confirming proper folding and activity through functional assays.
3. Biochemical Characterization and Stability Studies
This application is well-suited and should be prioritized. Techniques like circular dichroism spectroscopy, dynamic light scattering, and thermal stability assays can directly assess the protein's folding state, oligomerization, and stability. These studies are valuable even if the protein is inactive, as they characterize the recombinant hlgA itself and can inform about its suitability for other applications.
4. Tag-Based Detection Method Development
This application is appropriate and independent of folding status. The dual tags make this protein useful for developing and optimizing detection methods (e.g., Western blot, flow cytometry) using anti-His or anti-Myc antibodies. It can serve as a positive control for tag-based assays, but results may not reflect native hlgA behavior if the protein is misfolded.
Final Recommendation & Action Plan
Given that hlgA is a bacterial protein expressed in a homologous prokaryotic system, the likelihood of proper folding is high, but validation is essential. Recommend first performing functional validation (e.g., hemolytic assay with red blood cells) to confirm bioactivity, and biophysical characterization (e.g., size-exclusion chromatography for oligomeric state) to assess folding. Antibody development and tag-based detection applications can proceed immediately. Protein-protein interaction studies should await folding and activity validation. Always include appropriate controls, such as native hlgA or known binding partners, to ensure physiological relevance.
