Recombinant Neisseria meningitidis serogroup B Quinolinate synthase (nadA) is expressed in E. coli, spanning amino acids 1-370. It comes with an N-terminal 6xHis-tag that makes purification more straightforward. The protein appears to maintain purity levels above 85% based on SDS-PAGE analysis, which should provide reliable experimental results. This product is meant for research use only and contains minimal endotoxin levels, making it appropriate for various biochemical assays and studies.
The nadA gene encodes quinolinate synthase, an enzyme that participates in NAD (nicotinamide adenine dinucleotide) biosynthesis—a critical coenzyme for cellular metabolism. In Neisseria meningitidis, this protein likely plays a key role in synthesizing quinolinic acid, which serves as a precursor to NAD. Studying this enzyme may provide valuable insights into bacterial NAD biosynthesis, metabolic regulation, and potentially new antibiotic targets.
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 NAD Biosynthesis Pathway
This recombinant quinolinate synthase allows researchers to examine the de novo NAD biosynthesis pathway in Neisseria meningitidis through in vitro enzymatic assays. Scientists can explore the protein's catalytic properties, substrate specificity, and kinetic parameters with purified enzyme preparations. The N-terminal 6xHis tag helps with protein purification and immobilization for detailed biochemical work. Such research could advance our understanding of bacterial NAD metabolism and reveal species-specific differences in this essential biosynthetic pathway.
2. Antibody Development and Immunological Studies
The purified recombinant protein works well as an antigen for creating polyclonal or monoclonal antibodies against N. meningitidis quinolinate synthase. Researchers can develop these antibodies for various applications—Western blotting, immunofluorescence microscopy, and immunoprecipitation studies. The high purity level (>85%) should minimize cross-reactivity with other bacterial proteins during antibody production. These antibodies would become useful tools for studying protein expression, localization, and regulation in bacterial systems.
3. Protein-Protein Interaction Studies
The 6xHis-tagged quinolinate synthase can be used in pull-down assays to identify potential protein interaction partners within the NAD biosynthesis pathway or broader cellular networks. The tag allows efficient immobilization on nickel-affinity matrices for capturing interacting proteins from bacterial lysates. Co-immunoprecipitation experiments and yeast two-hybrid screens with this protein as bait might reveal novel regulatory mechanisms or metabolic pathway connections. These studies could help us understand how NAD biosynthesis integrates with other cellular processes in pathogenic bacteria.
4. Structural and Biophysical Analysis
This recombinant protein preparation appears suitable for structural biology applications including X-ray crystallography, NMR spectroscopy, and cryo-electron microscopy studies. The high purity and full-length expression (1-370aa) should provide good material for determining three-dimensional structure and conformational dynamics. Biophysical techniques like dynamic light scattering, analytical ultracentrifugation, and thermal stability assays can help characterize protein oligomerization states and stability parameters. Such structural insights might inform our understanding of enzyme mechanism and potential allosteric regulation sites.
