Recombinant Helicobacter pylori Bacterial non-heme ferritin (ftnA) is expressed in E. coli and contains the full-length protein spanning amino acids 1-167. The protein carries an N-terminal 10xHis-tag, which makes purification and detection more straightforward. SDS-PAGE analysis shows purity greater than 85%, suggesting this reagent should work well for most research applications.
The non-heme ferritin protein ftnA from Helicobacter pylori appears to play a key role in how these bacteria handle iron storage and detoxification - processes that seem critical for their survival and ability to cause disease. This protein is likely involved in iron metabolism pathways, where it captures excess iron to prevent cellular damage while keeping some available when supplies run low. Researchers studying bacterial iron balance and related cellular processes may find this protein particularly interesting.
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 Helicobacter pylori Bacterial non-heme ferritin (ftnA) is expressed in E. coli, a prokaryotic system that is generally suitable for expressing bacterial proteins like ftnA, as they share similar cellular environments and folding requirements. Ferritins are typically stable proteins that can often fold correctly in E. coli, especially when expressed in full-length (1-167aa). The N-terminal 10xHis tag may have minimal impact on folding, but since activity is explicitly unverified, the protein cannot be assumed to be correctly folded or bioactive without functional validation (e.g., iron-binding assays). The >85% purity indicates low contamination, but it does not confirm native conformation. Therefore, while the probability of correct folding is reasonably high, experimental confirmation is essential.
1. Protein-Protein Interaction Studies via His-Tag Pull-Down Assays
The N-terminal 10xHis-tag allows researchers to attach the recombinant H. pylori ferritin to nickel-based columns for pull-down experiments. This setup can help identify proteins or other molecules that might bind to bacterial non-heme ferritin in test tube conditions. Scientists can mix cell extracts or purified protein collections with the attached ferritin to trap any interacting partners, then use mass spectrometry to figure out what got caught. However, if the ferritin is misfolded, it may not interact physiologically with true binding partners, leading to non-specific or false interactions. The 85% purity level is sufficient, but results should be interpreted cautiously until folding and activity are validated.
2. Antibody Development and Immunoassay Applications
This recombinant H. pylori ferritin could work as an antigen for creating antibodies that specifically recognize this bacterial protein. The His-tagged version can be used in ELISA experiments to test how well antibodies bind and to measure their specificity. The purified protein may also serve as a positive control or reference standard in Western blots and similar detection methods. While the 85% purity might not be perfect, it appears adequate for immunization studies and antibody testing. Even if misfolded, antibodies may recognize linear epitopes, but validation against native ferritin is recommended for conformational epitopes.
3. Comparative Structural and Biochemical Analysis
Researchers can use this recombinant ferritin to compare bacterial non-heme ferritins with their eukaryotic counterparts through methods like dynamic light scattering, analytical ultracentrifugation, and various spectroscopic approaches. Since this represents the full-length protein (1-167aa), it should allow for meaningful structural comparisons with other ferritin family members. Scientists can also examine biochemical properties like heat stability, pH tolerance, and how the proteins cluster together - studies that might reveal what makes H. pylori ferritin unique. These studies are valuable even if the protein is misfolded, as they characterize the recombinant product. The His-tag helps standardize purification steps, which is important for consistent results.
4. Iron-Binding and Storage Mechanism Studies
This recombinant H. pylori ferritin can be used in laboratory tests that examine how well it binds and stores iron compared to other bacterial non-heme ferritins. Researchers can use light-based measurements and color-change assays to track iron uptake and release under different chemical conditions and pH levels. However, if the protein is misfolded, iron-binding activity may be impaired or absent, leading to inaccurate conclusions about H. pylori's iron handling. This application requires prior validation of iron-binding activity to ensure reliable results. The purified system minimizes interference, but folding must be confirmed.
Final Recommendation & Action Plan
Given the uncertainty in folding and bioactivity, it is recommended to first perform biochemical and functional validation to confirm the protein's conformation and iron-binding activity. Use techniques like size-exclusion chromatography (for oligomeric state), circular dichroism (for secondary structure), and iron-binding assays (e.g., using ferrozine or spectrophotometric methods) to assess functionality. If validated, the protein can be used for all described applications; if not, focus on antibody development and comparative biochemical studies, avoiding iron-binding and interaction studies. Always include appropriate controls, such as native ferritin or known iron-binding standards, and consider using activity-verified proteins for critical functional assays. The E. coli expression system is appropriate for this bacterial protein, but verification remains crucial.
