Recombinant Escherichia coli dITP/XTP pyrophosphatase (rdgB) is expressed in E. coli and covers the full length of the protein (1-197 amino acids). This product includes an N-terminal 10xHis-SUMO tag and a C-terminal Myc tag, which help with purification and detection. The protein appears to be purified to over 85% purity, as determined by SDS-PAGE, suggesting it should work well for research applications.
dITP/XTP pyrophosphatase is an essential enzyme in Escherichia coli that's involved in nucleotide metabolism. It breaks down non-canonical nucleotides like dITP and XTP to prevent their incorporation into DNA, which helps maintain genomic stability. This function makes it an important subject for studying DNA replication fidelity and how cells respond to nucleotide pool imbalances.
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 Tag-Assisted Pull-Down Assays
The dual-tagged nature of this recombinant rdgB protein, with both N-terminal His-SUMO and C-terminal Myc tags, seems well-suited for investigating protein-protein interactions within nucleotide metabolism pathways. The His tag can be used for immobilization on nickel-affinity matrices, while the Myc tag allows detection and validation of binding partners through Western blot analysis. This approach might help identify novel interacting proteins involved in DNA repair and replication processes where dITP/XTP pyrophosphatase activity is relevant. The 85% purity level appears sufficient for pull-down experiments where specific interactions can be distinguished from non-specific binding.
2. Antibody Development and Validation Platform
The recombinant rdgB protein can work as an antigen for generating specific antibodies against E. coli dITP/XTP pyrophosphatase. Both His-SUMO and Myc tags provide built-in controls for antibody specificity testing, allowing researchers to distinguish between antibodies that recognize the target protein versus those that cross-react with the tags. The protein's purity level of greater than 85% should be adequate for immunization protocols and subsequent antibody characterization through ELISA, Western blot, and other immunoassays. This would be particularly valuable for researchers studying nucleotide pool sanitization mechanisms in bacterial systems.
3. Biochemical Characterization and Substrate Specificity Analysis
This recombinant protein provides a useful tool for detailed biochemical characterization of E. coli rdgB enzyme properties, including substrate binding affinity, cofactor requirements, and optimal reaction conditions. Researchers can investigate the protein's interaction with various nucleotide substrates and analogs to better understand its role in maintaining nucleotide pool integrity. The dual tagging system makes protein purification and detection easier throughout biochemical assays, while the 85% purity is likely sufficient for most enzymatic characterization studies. Such studies would contribute to understanding bacterial DNA repair mechanisms and nucleotide metabolism pathways.
4. Structural and Biophysical Studies
The recombinant rdgB protein can be used in various biophysical analyses to investigate its structural properties and conformational dynamics. Techniques such as dynamic light scattering, circular dichroism spectroscopy, and analytical ultracentrifugation can provide insights into protein folding, stability, and oligomerization states. The His tag makes consistent protein purification easier for reproducible biophysical measurements, while the known molecular weight and composition allow for accurate data interpretation. These studies would enhance understanding of the structure-function relationships in this class of pyrophosphatases and inform comparative analyses with homologous enzymes from other species.
