Recombinant Mouse Pyridoxal phosphate phosphatase (Pdxp) is produced in a yeast system, which appears to provide efficient and reliable results. The full-length protein covers amino acids 1 to 292 and includes an N-terminal 6xHis tag that makes purification and detection more straightforward. SDS-PAGE analysis confirms purity levels above 90%, though this protein is intended specifically for research applications where high-quality reagents matter most.
Pyridoxal phosphate phosphatase (Pdxp) is an enzyme that removes phosphate groups from pyridoxal 5'-phosphate (PLP), the active form of vitamin B6. It likely plays an important role in controlling PLP levels, which are essential for many metabolic pathways. Research on Pdxp may reveal new insights into how metabolic processes are regulated and how enzymes function, making it a useful tool for biochemistry and physiology studies.
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.
Based on the provided information, recombinant mouse Pdxp is produced in a yeast expression system as a full-length protein (1-292aa) with an N-terminal 6xHis-tag. Yeast expression systems provide eukaryotic folding machinery capable of supporting proper disulfide bond formation and post-translational modifications, which are important for phosphatase activity and stability. As a full-length protein expressed in a eukaryotic system, it has a high probability of correct folding. However, phosphatase activity requires precise active site formation and potential metal cofactor incorporation. The N-terminal His-tag is relatively small and unlikely to significantly interfere with folding or function. No validation data (e.g., phosphatase activity assays, circular dichroism) are provided. Therefore, while the protein is likely correctly folded, its bioactivity cannot be confirmed without experimental validation.
1. Biochemical Characterization of Pyridoxal Phosphate Phosphatase Activity
If the recombinant Pdxp is correctly folded and functional, it can be used for enzyme kinetics studies to characterize substrate specificity and kinetic parameters. However, if inactive (due to improper folding or lack of cofactors), activity assays would yield invalid results. The His-tag is unlikely to interfere with the active site, but validation of enzymatic activity is essential before quantitative kinetic studies.
2. Antibody Development and Validation Studies
This application is highly suitable as antibody generation primarily relies on linear epitope recognition. The full-length protein provides comprehensive epitope coverage, and the high purity ensures minimal cross-reactivity. Antibodies generated will likely recognize both linear and conformational epitopes of native Pdxp.
3. Protein-Protein Interaction Studies
If properly folded, the His-tagged Pdxp can be used in pull-down assays to identify interaction partners in vitamin B6 metabolic pathways. However, validation of folding is recommended as misfolding could alter interaction interfaces. The tag may cause steric hindrance in some interactions, but generally allows effective immobilization.
4. Structural and Biophysical Analysis
If correctly folded, the protein is suitable for structural studies. The His-tag should be removed for high-resolution structural determination (X-ray crystallography, NMR) but may not significantly interfere with biophysical characterization (circular dichroism, thermal stability). Misfolding would compromise structural data relevance.
5. Comparative Species Analysis
If functionally active, the recombinant Pdxp enables valid comparative studies with orthologs from other species. However, activity validation is essential first, as improper folding would make cross-species comparisons biologically meaningless. The yeast expression system likely produces proteins with correct folding for meaningful evolutionary analysis.
Final Recommendation & Action Plan
This yeast-expressed full-length Pdxp has a high probability of being properly folded and functional due to the eukaryotic expression system and full-length construct. The recommended action plan includes: (1) First validate enzymatic activity using pyridoxal phosphate as substrate with appropriate controls; (2) Confirm proper folding through biophysical methods (circular dichroism for secondary structure, size-exclusion chromatography for oligomeric state); (3) Proceed with proposed applications once activity is confirmed - antibody development can begin immediately while functional studies should await activity validation; (4) For structural studies, consider removing the His-tag proteolytically for highest resolution analysis; (5) Always include positive controls (known active phosphatase) and negative controls in experiments. The protein can be used with high confidence for most applications after basic activity validation.
Recombinant Mouse Pyridoxal phosphate phosphatase (Pdxp) is produced in a yeast system, which appears to provide efficient and reliable results. The full-length protein covers amino acids 1 to 292 and includes an N-terminal 6xHis tag that makes purification and detection more straightforward. SDS-PAGE analysis confirms purity levels above 90%, though this protein is intended specifically for research applications where high-quality reagents matter most.
Pyridoxal phosphate phosphatase (Pdxp) is an enzyme that removes phosphate groups from pyridoxal 5'-phosphate (PLP), the active form of vitamin B6. It likely plays an important role in controlling PLP levels, which are essential for many metabolic pathways. Research on Pdxp may reveal new insights into how metabolic processes are regulated and how enzymes function, making it a useful tool for biochemistry and physiology studies.
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.
Based on the provided information, recombinant mouse Pdxp is produced in a yeast expression system as a full-length protein (1-292aa) with an N-terminal 6xHis-tag. Yeast expression systems provide eukaryotic folding machinery capable of supporting proper disulfide bond formation and post-translational modifications, which are important for phosphatase activity and stability. As a full-length protein expressed in a eukaryotic system, it has a high probability of correct folding. However, phosphatase activity requires precise active site formation and potential metal cofactor incorporation. The N-terminal His-tag is relatively small and unlikely to significantly interfere with folding or function. No validation data (e.g., phosphatase activity assays, circular dichroism) are provided. Therefore, while the protein is likely correctly folded, its bioactivity cannot be confirmed without experimental validation.
1. Biochemical Characterization of Pyridoxal Phosphate Phosphatase Activity
If the recombinant Pdxp is correctly folded and functional, it can be used for enzyme kinetics studies to characterize substrate specificity and kinetic parameters. However, if inactive (due to improper folding or lack of cofactors), activity assays would yield invalid results. The His-tag is unlikely to interfere with the active site, but validation of enzymatic activity is essential before quantitative kinetic studies.
2. Antibody Development and Validation Studies
This application is highly suitable as antibody generation primarily relies on linear epitope recognition. The full-length protein provides comprehensive epitope coverage, and the high purity ensures minimal cross-reactivity. Antibodies generated will likely recognize both linear and conformational epitopes of native Pdxp.
3. Protein-Protein Interaction Studies
If properly folded, the His-tagged Pdxp can be used in pull-down assays to identify interaction partners in vitamin B6 metabolic pathways. However, validation of folding is recommended as misfolding could alter interaction interfaces. The tag may cause steric hindrance in some interactions, but generally allows effective immobilization.
4. Structural and Biophysical Analysis
If correctly folded, the protein is suitable for structural studies. The His-tag should be removed for high-resolution structural determination (X-ray crystallography, NMR) but may not significantly interfere with biophysical characterization (circular dichroism, thermal stability). Misfolding would compromise structural data relevance.
5. Comparative Species Analysis
If functionally active, the recombinant Pdxp enables valid comparative studies with orthologs from other species. However, activity validation is essential first, as improper folding would make cross-species comparisons biologically meaningless. The yeast expression system likely produces proteins with correct folding for meaningful evolutionary analysis.
Final Recommendation & Action Plan
This yeast-expressed full-length Pdxp has a high probability of being properly folded and functional due to the eukaryotic expression system and full-length construct. The recommended action plan includes: (1) First validate enzymatic activity using pyridoxal phosphate as substrate with appropriate controls; (2) Confirm proper folding through biophysical methods (circular dichroism for secondary structure, size-exclusion chromatography for oligomeric state); (3) Proceed with proposed applications once activity is confirmed - antibody development can begin immediately while functional studies should await activity validation; (4) For structural studies, consider removing the His-tag proteolytically for highest resolution analysis; (5) Always include positive controls (known active phosphatase) and negative controls in experiments. The protein can be used with high confidence for most applications after basic activity validation.
