Recombinant Escherichia coli O157:H7 Translocated intimin receptor Tir is expressed in E. coli, covering the extracellular domain region of 252-362 amino acids. The protein carries an N-terminal 6xHis-SUMO tag to help with purification and reaches purity levels above 90%, as confirmed through SDS-PAGE analysis. This product is made specifically for research purposes and is not intended for clinical applications.
The Translocated intimin receptor (Tir) appears to be a critical component in how Escherichia coli O157:H7 causes disease, particularly in its interactions with host cells. When Tir gets translocated into host cells, it seems to integrate into the host membrane, where it likely functions as a receptor for the bacterial adhesin intimin. This interaction may be essential for forming attaching and effacing lesions, which makes Tir an important target for researchers studying bacterial pathogenesis and how pathogens 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.
1. Antibody Development and Characterization Studies
This recombinant Tir extracellular domain can work as an immunogen for creating polyclonal or monoclonal antibodies that specifically target E. coli O157:H7. The purified protein offers a well-defined antigen for immunization protocols and later antibody screening assays. The N-terminal His-SUMO tag makes purification and immobilization straightforward for ELISA-based antibody characterization. Scientists can work with this protein to develop research-grade antibodies for investigating E. coli O157:H7 pathogenesis mechanisms.
2. Protein-Protein Interaction Studies
Researchers might use the extracellular domain of Tir in pull-down assays to identify and characterize host cell surface receptors or other bacterial proteins that bind to this region. The His-tag allows for immobilization on nickel-affinity matrices, which can capture potential binding partners from cell lysates or purified protein libraries. Surface plasmon resonance or other biophysical techniques could work with this recombinant protein to measure binding kinetics and affinities with suspected interaction partners.
3. Structural and Biochemical Characterization
This purified Tir extracellular domain supplies material for structural biology studies, including X-ray crystallography, NMR spectroscopy, or cryo-electron microscopy. Its high purity level appears suitable for biophysical characterization techniques like circular dichroism spectroscopy to analyze secondary structure content. The defined expression region (252-362aa) may offer a stable domain for investigating the molecular architecture of this bacterial virulence factor.
4. In Vitro Binding and Competition Assays
Scientists can work with the recombinant protein to develop cell-free binding assays that study the molecular mechanisms behind E. coli O157:H7 adhesion. Researchers might use this protein in competition experiments to screen for small molecules or peptides that disrupt Tir-mediated interactions. If needed, the SUMO tag can be removed enzymatically to study the native extracellular domain without potential tag interference in sensitive binding studies.
