Recombinant Human Selenide, water dikinase 1 (SEPHS1) is expressed in E. coli and covers the complete sequence from amino acids 1 to 392. The protein carries an N-terminal GST tag, which appears to make purification simpler while potentially improving stability. SDS-PAGE analysis confirms purity levels above 90%. This material seems well-suited for research applications, though results may vary depending on specific experimental conditions.
Selenide, water dikinase 1 (SEPHS1) likely plays a central role in selenoprotein biosynthesis through its involvement in converting selenide to selenophosphate. The enzyme appears essential for selenium metabolism, a process that may be critical for various biological pathways. Studying this protein could provide insights into how selenium affects cellular processes and its broader role in human health, though much remains to be understood about these mechanisms.
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.
Human SEPHS1 is an enzyme that catalyzes the conversion of selenide to selenium phosphate, requiring ATP and magnesium as cofactors. Its activity depends on correct folding to form the active site. The E. coli expression system can produce soluble proteins, and the GST tag may enhance solubility. However, enzyme activity requires precise tertiary structure and cofactor binding. While the full-length protein (1-392aa) contains all functional domains, the GST tag might sterically affect the active site or folding. The probability of correct folding is moderate, but enzymatic activity must be experimentally verified.
1. GST Pull-Down Assays for Protein-Protein Interaction Studies
This application is feasible but carries a moderate risk. Protein-protein interactions depend on native conformation. If correctly folded, SEPHS1 could identify physiological interaction partners in selenium metabolism pathways. However, the GST tag may sterically hinder interactions or cause non-specific binding. A misfolded protein may yield false positives or negatives. Results require validation with an endogenous protein. The GST tag and potential misfolding pose risks, but the full-length design supports interaction studies if activity is confirmed.
2. Antibody Development and Validation
This recombinant SEPHS1 is an excellent immunogen for generating antibodies against human SEPHS1. The full-length sequence ensures broad epitope coverage. The GST tag facilitates purification and screening. Antibodies may recognize both linear and conformational epitopes if the protein is properly folded.
3. Biochemical Characterization and Enzyme Kinetics Studies
This is the critical first step to assess protein quality and activity. Techniques like SEC-MALS can determine oligomeric state, while enzyme assays with ATP and selenide can verify catalytic activity. Stability studies (thermal shift assays) can optimize conditions for functional use.
4. GST-Based ELISA Development
This protein is well-suited as a standard for quantitative ELISA to measure SEPHS1 immunoreactivity. The GST tag enables consistent immobilization. Functional ELISAs (e.g., detecting enzyme activity) require activity confirmation.
Final Recommendation & Action Plan
This recombinant SEPHS1 has potential for functional applications but requires activity validation before reliable use in interaction or kinetic studies. The immediate priority is Application 3 (Biochemical Characterization) to confirm enzyme activity with ATP/selenide substrates and assess folding quality. If active, it becomes suitable for Applications 1 and the functional aspects of Application 4. Application 2 (Antibody Development) can proceed immediately. For all functional uses, verify activity first. If inactive, limit use to Applications 2 and 4 as an immunoassay standard. This systematic approach ensures reliable outcomes.
