Recombinant Mycobacterium tuberculosis Mycothione reductase (mtr) gets produced in E. coli and contains the complete protein spanning amino acids 1-459. The protein carries a 10xHis-SUMO tag at the N-terminus and a Myc tag at the C-terminus, which helps with purification and detection. SDS-PAGE analysis shows the purity exceeds 85%, making it suitable for research work.
Mycothione reductase appears to be a key enzyme in the antioxidant defense system of Mycobacterium tuberculosis. It seems to play an important role in maintaining redox balance by reducing mycothiol disulfide back to mycothiol. This function may be vital for protecting the bacterium from oxidative stress - something that's likely crucial for its survival and ability to cause disease. Given this role, it has become a significant focus in tuberculosis 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. Antibody Development and Immunoassay Studies
The dual-tagged recombinant mycothione reductase can work as an immunogen for creating polyclonal or monoclonal antibodies that target the M. tuberculosis mtr protein. Both the N-terminal His-SUMO and C-terminal Myc tags offer multiple recognition sites for antibodies and allow validation through tag-specific detection methods. These antibodies could prove valuable for studying how mycothione reductase gets expressed, where it localizes, and how it's regulated in mycobacterial research. The >85% purity appears sufficient for immunization protocols and follow-up antibody characterization work.
2. Protein-Protein Interaction Studies Using Tag-Assisted Pull-Down Assays
That N-terminal His tag allows for nickel-affinity based pull-down experiments to hunt for potential binding partners of mycothione reductase in M. tuberculosis protein extracts or recombinant protein libraries. Meanwhile, the C-terminal Myc tag can be used for immunoprecipitation studies with anti-Myc antibodies to confirm interactions found through His-tag pull-downs. These two tagging approaches may provide solid methods for mapping out the protein interaction network of mycothione reductase in mycobacterial metabolism research.
3. Biochemical Characterization and Enzyme Kinetics Analysis
The full-length recombinant protein (1-459aa) offers a complete molecular framework for investigating mycothione reductase's biochemical properties. This includes protein stability, cofactor binding, and substrate specificity studies. While biological activity hasn't been tested yet, the protein could be put through various biochemical assays to figure out optimal conditions for potential enzymatic activity. The dual tags make protein purification and detection easier throughout these characterization experiments.
4. Comparative Protein Structure and Function Studies
The recombinant mtr protein might be useful in comparative studies with mycothione reductases from other mycobacterial strains or related oxidoreductases to understand how structure relates to function. The >85% purity level should support biophysical characterization techniques like dynamic light scattering, circular dichroism spectroscopy, and thermal stability assays. These studies could provide insights into protein folding, stability, and conformational changes under different experimental conditions that are relevant to mycobacterial physiology research.
