Recombinant Escherichia coli Lactose permease (lacY) is produced using an in vitro E.coli expression system, featuring an N-terminal 10xHis-tag for simplified purification. This partial protein covers amino acids 1-250 and achieves a purity greater than 85% as confirmed by SDS-PAGE. It's designed for research use only, with no endotoxin level specified, ensuring it meets the quality standards necessary for laboratory applications.
Lactose permease appears to be a critical component in Escherichia coli, helping transport lactose across the cell membrane. As a symporter, it plays what seems to be a fundamental role in sugar metabolism by coupling lactose transport with proton translocation. This enables efficient lactose uptake. Studying this protein may be pivotal in understanding membrane transport mechanisms and could provide insights into similar processes in other organisms.
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
This N-terminal His-tagged lacY fragment can be used in pull-down assays to identify and characterize proteins that interact with lactose permease. The His-tag allows immobilization on nickel-affinity resins, letting researchers capture potential binding partners from E. coli cell lysates or purified protein libraries. Such studies might reveal regulatory proteins, chaperones, or other membrane-associated factors that could modulate lactose permease function or assembly.
2. Antibody Development and Validation
The recombinant lacY fragment serves as an excellent antigen for generating specific antibodies against lactose permease. Its high purity (>85%) and defined amino acid sequence (1-250aa) make it suitable for immunizing laboratory animals or screening existing antibody collections. Researchers can use this protein in ELISA-based assays to validate antibody specificity and determine binding affinities for subsequent immunoblotting or immunofluorescence applications.
3. Structural and Biochemical Characterization
This partial lacY protein can be used for biophysical studies to understand the structural properties of the N-terminal region of lactose permease. Techniques such as circular dichroism spectroscopy, dynamic light scattering, or limited proteolysis experiments may provide insights into protein folding, stability, and domain organization. The His-tag makes purification easier for these analytical methods while maintaining protein integrity.
4. In Vitro Binding Assays
The recombinant lacY fragment can be used in direct binding studies to investigate interactions with small molecules, substrates, or inhibitors that may associate with the N-terminal region of lactose permease. The His-tag allows easy immobilization for surface plasmon resonance or other label-free binding detection methods. These assays can help map binding sites and determine binding kinetics for compounds of interest in lactose transport research.
