Recombinant Escherichia coli Lipopolysaccharide export system protein lptA is produced using an E. coli expression system. The protein includes the complete mature sequence spanning residues 28 to 185. For easier purification and detection, it carries an N-terminal 6xHis-SUMO tag. SDS-PAGE analysis shows the protein reaches over 90% purity, which should provide high-quality material for research work.
LptA appears to be a key component in E. coli's lipopolysaccharide (LPS) transport machinery. The protein likely plays an important role in assembling and moving LPS molecules across the bacterial cell envelope—a process that seems critical for outer membrane integrity and function. This makes the protein particularly useful for researchers studying bacterial membrane biology and LPS transport 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.
1. Protein-Protein Interaction Studies Using Pull-Down Assays
The N-terminal 6xHis-SUMO tag allows for nickel affinity-based pull-down experiments. These can help identify possible binding partners of LptA within the lipopolysaccharide transport pathway. Researchers can attach the recombinant protein to nickel-coated beads, then mix it with E. coli cell lysates or specific candidate proteins. Any co-precipitated proteins can then be examined through mass spectrometry or Western blotting to map out interaction networks. The high purity level (>90%) should reduce background binding from contaminating proteins during these studies.
2. Antibody Development and Validation
This purified recombinant LptA protein works well as an antigen for creating polyclonal or monoclonal antibodies that target the mature LptA protein. The combination of high purity (>90%) and proper folding of the mature protein region (28-185aa) expressed in E. coli may provide authentic epitopes for immunization. Scientists can validate resulting antibodies using ELISA with the His-tagged protein coated on plates. Specificity testing through Western blotting against the recombinant protein offers another validation approach.
3. Biochemical Characterization and Stability Studies
The purified LptA protein enables detailed biochemical analysis. This might include thermal stability assays, pH tolerance studies, and buffer optimization experiments. Differential scanning fluorimetry or circular dichroism spectroscopy can evaluate protein folding and stability across different conditions. The SUMO tag appears to boost protein solubility and stability, potentially making it well-suited for extended biochemical studies aimed at understanding the protein's biophysical properties.
4. In Vitro Reconstitution Assays for LPS Transport Studies
This recombinant LptA can be added to in vitro reconstitution experiments designed to study lipopolysaccharide transport mechanisms. When combined with other purified Lpt pathway components, it may help researchers understand the transport process better. The protein works in liposome-based assays or proteoliposome reconstitution studies to examine its role in LPS movement. The His-tag makes purification straightforward and allows for easy concentration adjustments needed for stoichiometric reconstitution experiments with other pathway proteins.
