Recombinant Human Glutathione S-transferase P (GSTP1) is expressed in E. coli and comprises the full length of the mature protein (amino acids 2-210). It features an N-terminal 6xHis tag for ease of purification and detection. The protein is purified to greater than 90% purity, as confirmed by SDS-PAGE analysis, ensuring high-quality standards for research applications.
GSTP1 belongs to the glutathione S-transferase family and appears to play a crucial role in cellular detoxification processes. The enzyme catalyzes the conjugation of glutathione to various electrophilic compounds, which may help with xenobiotic metabolism and protection against oxidative stress. Researchers often examine GSTP1 for its role in cellular defense mechanisms, though its broader implications in different research contexts continue to emerge.
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.
The human GSTP1 is a soluble enzyme that typically folds well in E. coli, as it does not require complex post-translational modifications like glycosylation. The small His tag is unlikely to significantly interfere with folding, and the expression of the mature protein sequence suggests all functional domains are present. E. coli is a common and effective system for expressing active GST proteins. Therefore, there is a high probability that this protein is correctly folded and bioactive. However, experimental validation (e.g., enzyme activity assays) is recommended to confirm functionality before use in sensitive applications.
1. Glutathione Conjugation Assays and Enzyme Kinetics Studies
If the recombinant GSTP1 is verified to be enzymatically active (e.g., through glutathione conjugation assays), it can be used to investigate catalytic properties and determine kinetic parameters (Km, Vmax) with various substrates. The high purity supports reliable results, and the His tag facilitates purification and immobilization. However, without activity confirmation, results may be misleading due to potential misfolding.
2. Protein-Protein Interaction Studies
Provided that the protein is correctly folded, it may be used in pull-down assays to identify binding partners, leveraging the His tag for immobilization. However, misfolding could alter interaction sites, leading to non-specific binding. Validation with native GSTP1 from eukaryotic sources is advised for physiological relevance.
3. Antibody Development and Validation
This recombinant GSTP1 is suitable as an antigen for antibody production, as antibodies can recognize linear epitopes even if the protein is partially misfolded. The high purity minimizes background in immunization and screening. However, antibodies should be validated against native GSTP1 to ensure recognition of conformational epitopes in applications like Western blot or ELISA.
4. Structural Biology and Biophysical Characterization
If the protein is correctly folded, it is appropriate for structural studies (e.g., X-ray crystallography) and biophysical analyses (e.g., circular dichroism, dynamic light scattering). The His tag can be removed if necessary. However, a misfolded protein would yield inaccurate structural insights, so folding should be confirmed first.
5. Inhibitor Screening and Drug Discovery Research
This protein can be used for inhibitor screening only if enzymatically active. High-throughput assays require validated activity to ensure reliable IC50 determinations. Without confirmation, screening results may be invalid due to potential lack of enzyme function.
Final Recommendation & Action Plan
Before using this recombinant GSTP1 protein for any functional application, prioritize experimental validation of its folding and bioactivity. Start with a simple glutathione conjugation assay using a standard substrate (e.g., CDNB) to confirm enzyme activity and measure kinetic parameters. If active, proceed with intended studies; if not, investigate folding issues via biophysical methods (e.g., circular dichroism). For applications like antibody production, proceed with caution and validate antibodies against native protein. The high purity and mature sequence suggest good potential, but verification is key to ensure reliable outcomes.
