Recombinant Klebsiella pneumoniae Undecaprenyl phosphate-alpha-4-amino-4-deoxy-L-arabinose arabinosyl transferase (arnT) is expressed in E. coli and includes an N-terminal 6xHis-tag for simplified purification. The product represents a partial protein spanning amino acids 429-551 and achieves greater than 90% purity as determined by SDS-PAGE. This research-use-only protein serves as a reliable tool for laboratory studies requiring high-quality reagents.
The arnT gene in Klebsiella pneumoniae encodes a protein that appears to play a crucial role in bacterial resistance mechanisms. Acting as an arabinosyl transferase, it modifies lipopolysaccharides—key components of the bacterial outer membrane. This modification may influence the bacterium's susceptibility to certain antibiotics, which makes arnT a significant focus in microbial resistance studies and a potentially valuable target for antibacterial research.
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. Biochemical Characterization of ArnT Enzymatic Properties
Researchers can use this recombinant ArnT fragment (429-551aa) to investigate biochemical properties of the C-terminal region of undecaprenyl phosphate-alpha-4-amino-4-deoxy-L-arabinose arabinosyl transferase from Klebsiella pneumoniae. The high purity (>90%) likely makes it suitable for kinetic studies, substrate binding assays, and cofactor requirement analysis to understand catalytic mechanisms. The N-terminal His-tag simplifies purification and immobilization for enzyme activity measurements. This partial protein may retain specific functional domains that contribute to overall transferase activity, though further validation would be necessary.
2. Antibody Development and Validation
The purified recombinant ArnT fragment serves as a potentially excellent immunogen for generating polyclonal or monoclonal antibodies specific to Klebsiella pneumoniae ArnT. Researchers can use the His-tagged protein in ELISA-based screening assays to identify and characterize antibody specificity and affinity. These antibodies would become valuable research tools for studying ArnT expression, localization, and regulation in bacterial cells. High purity should ensure minimal cross-reactivity with other bacterial proteins during antibody development.
3. Protein-Protein Interaction Studies
Scientists may use the His-tagged ArnT fragment in pull-down assays to identify potential protein partners involved in lipopolysaccharide modification pathways in Klebsiella pneumoniae. The tag allows immobilization on nickel-affinity matrices for capturing interacting proteins from bacterial lysates. This approach could reveal regulatory proteins or other enzymes that form complexes with ArnT during arabinose transfer reactions. Such studies might advance understanding of molecular networks controlling bacterial cell wall biosynthesis.
4. Structural and Biophysical Analysis
This recombinant protein fragment provides material for structural biology studies including X-ray crystallography, NMR spectroscopy, or cryo-electron microscopy to elucidate the three-dimensional structure of the ArnT C-terminal region. High purity is essential for obtaining quality crystals or NMR spectra. Biophysical techniques such as dynamic light scattering, circular dichroism, and thermal stability assays can characterize protein folding, stability, and conformational changes. These studies would likely provide insights into structure-function relationships within the arabinosyl transferase family.
5. Inhibitor Screening and Drug Discovery Research
The purified ArnT fragment can serve as a target for high-throughput screening of small molecule libraries to identify potential inhibitors of arabinose transfer activity. The His-tag makes protein immobilization in microplate-based assays more straightforward for compound screening. Identified hits can be further characterized for binding affinity, specificity, and mechanism of inhibition using biochemical assays. This research application supports development of new antimicrobial strategies targeting bacterial cell wall modification enzymes, though clinical translation would require extensive additional work.
