Recombinant Staphylococcus epidermidis Endoribonuclease MazF is produced in E. coli and includes an N-terminal 6xHis-tag that makes purification and detection more straightforward. The protein appears to be expressed as a complete construct covering amino acids 1-120. SDS-PAGE analysis confirms purity levels exceeding 90%, which suggests this product may be well-suited for research applications requiring high-quality recombinant proteins.
Endoribonuclease MazF is recognized for its RNA cleavage activity—it seems to recognize and cut RNA at particular sequences. Bacterial stress responses likely involve this enzyme, and researchers have been examining its role in pathways that regulate cell growth. MazF's capacity to target RNA makes it potentially valuable for studies focused on gene expression control and how cells respond to stress.
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 His-Tag Affinity Purification
The N-terminal 6xHis-tag allows researchers to use nickel-based affinity purification methods for identifying possible binding partners or regulatory proteins that might interact with MazF endoribonuclease. Pull-down experiments could work with bacterial lysates or purified protein collections to capture proteins that appear to associate specifically with MazF. The >90% purity should minimize background interference from contaminating proteins during these interaction studies. This strategy might uncover components of toxin-antitoxin regulatory networks or cellular factors that could modulate MazF function.
2. Biochemical Characterization and Enzyme Kinetics Analysis
Researchers can use the purified recombinant MazF protein to develop in vitro ribonuclease activity assays with synthetic RNA substrates or bacterial RNA extracts. Optimal reaction conditions—pH, temperature, salt concentration, and cofactor requirements—can be determined through systematic testing. The high purity level should allow for precise protein quantification and what appears to be reliable kinetic parameter measurements. These studies would likely provide fundamental biochemical data on how S. epidermidis MazF endoribonuclease functions catalytically.
3. Antibody Development and Immunological Applications
The recombinant protein may serve as an effective immunogen for producing polyclonal or monoclonal antibodies specific to S. epidermidis MazF. The >90% purity should reduce cross-reactivity with other bacterial proteins during immunization procedures. Resulting antibodies could be useful for Western blotting, immunoprecipitation, or immunofluorescence studies to detect native MazF expression in bacterial cultures. The His-tag also makes it possible to develop sandwich ELISA assays for quantitative detection work.
4. Comparative Structure-Function Studies
The full-length recombinant protein (1-120aa) provides material for comparative analysis with MazF variants from other bacterial species. Proteolytic digestion experiments might map functional domains and identify regions that appear critical for ribonuclease activity. The purified protein enables systematic mutagenesis studies where researchers can modify specific amino acid residues and assess their effects on protein stability or function. Cross-species activity comparisons using standardized RNA substrates may reveal evolutionary conservation of catalytic mechanisms—though this remains to be fully established.
