Recombinant Escherichia coli 3-demethoxyubiquinol 3-hydroxylase (ubiF) is expressed in E.coli and includes an N-terminal 6xHis-tag that makes purification more straightforward. The product contains the full-length protein from amino acids 1 to 391, reaching a purity greater than 85% as verified by SDS-PAGE. This protein is designed solely for research purposes and appears to be a reliable tool for scientific investigation.
3-demethoxyubiquinol 3-hydroxylase is an enzyme that participates in the biosynthesis of ubiquinone, also known as coenzyme Q. This protein plays what seems to be a crucial role in the electron transport chain, helping to drive cellular energy production. Its activity is likely essential for maintaining cellular respiration and metabolic efficiency, which makes it an important subject of study in microbial and biochemical 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. Ubiquinone Biosynthesis Pathway Studies
This recombinant ubiF protein may serve as a valuable tool for investigating the ubiquinone biosynthesis pathway in E. coli through in vitro biochemical assays. Researchers can potentially use this protein to study the enzymatic conversion of 3-demethoxyubiquinol to ubiquinol-8, which could help clarify the molecular mechanisms of this critical step in coenzyme Q biosynthesis. The full-length protein (1-391aa) provides what appears to be the complete functional domain architecture necessary for comprehensive structure-function relationship studies. Such research might contribute to understanding how bacteria synthesize this essential electron transport chain component.
2. Protein-Protein Interaction Studies
The N-terminal 6xHis tag allows for pull-down assays to identify potential protein partners that interact with ubiF in the ubiquinone biosynthesis machinery. This approach could help map the protein interaction network involved in coenzyme Q production and identify regulatory proteins or enzyme complexes. The high purity (>85%) suggests reliable results in co-immunoprecipitation experiments and other interaction-based assays. These studies might reveal how ubiF integrates with other biosynthetic enzymes and cellular regulatory mechanisms.
3. Antibody Development and Validation
This purified recombinant protein can work as an immunogen for generating specific antibodies against E. coli ubiF, or as a standard for validating existing antibodies. The high purity and full-length nature of the protein make it suitable for immunization protocols and subsequent antibody characterization through ELISA, Western blotting, and other immunoassays. Such antibodies would likely be valuable research tools for studying ubiF expression levels, cellular localization, and regulation in various experimental conditions.
4. Comparative Enzymology and Evolution Studies
This recombinant ubiF protein can be used in comparative studies with homologous enzymes from other bacterial species to understand evolutionary relationships and functional conservation within the ubiquinone biosynthesis pathway. Researchers can perform side-by-side biochemical comparisons to identify conserved and divergent functional properties across different bacterial lineages. The standardized expression system and purification approach appears to make reproducible comparative analyses possible, which may inform our understanding of how this essential metabolic pathway has evolved.
