Recombinant Human Valacyclovir hydrolase (BPHL) is produced in a yeast expression system, spanning the full length of the mature protein from amino acids 38 to 291. The protein comes with an N-terminal 10xHis-tag and a C-terminal Myc-tag, which makes purification and detection more straightforward. SDS-PAGE analysis indicates purity levels above 90%, suggesting it may be suitable for various research applications.
Valacyclovir hydrolase, also known as BPHL, appears to play a critical role in activating antiviral prodrugs. This enzyme handles the hydrolysis of valacyclovir to its active form, acyclovir—a conversion that seems crucial for therapeutic efficacy. BPHL participates in drug metabolism pathways and has drawn considerable interest in pharmacological research, particularly for optimizing antiviral treatments.
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. Enzyme Kinetics and Substrate Specificity Studies
This recombinant BPHL protein can help characterize the enzymatic properties of valacyclovir hydrolase through in vitro biochemical assays. Researchers might determine kinetic parameters like Km, Vmax, and kcat using valacyclovir and other potential substrates under controlled reaction conditions. The high purity (>90%) should provide reliable quantitative measurements of enzyme activity. These studies could offer fundamental insights into the catalytic mechanism and substrate preferences of this human enzyme.
2. Protein-Protein Interaction Studies
The dual-tagged design of this protein (N-terminal His-tag and C-terminal Myc-tag) makes it potentially useful for investigating protein interactions through pull-down assays and co-immunoprecipitation experiments. The His-tag allows immobilization on nickel-based resins for capturing interacting partners from cell lysates or purified protein mixtures. Meanwhile, the Myc-tag provides another option for detection and validation of interactions using anti-Myc antibodies in Western blotting or immunofluorescence applications.
3. Antibody Development and Validation
This purified recombinant protein serves as a promising antigen for generating specific antibodies against human BPHL. The high purity level suggests that resulting antibodies will likely have minimal cross-reactivity with contaminants. Researchers can use the protein for immunizing animals, screening hybridomas, and validating antibody specificity through ELISA, Western blotting, and other immunoassays—with the incorporated Myc-tag acting as a positive control.
4. Structural and Biophysical Characterization
The recombinant BPHL protein appears suitable for biophysical studies aimed at understanding its structural properties and stability. Techniques like circular dichroism spectroscopy, dynamic light scattering, and thermal stability assays may provide insights into protein folding, secondary structure content, and conformational changes under different conditions. The yeast expression system typically produces properly folded eukaryotic proteins, which makes this preparation likely appropriate for structural studies.
5. Drug Metabolism Research Models
This recombinant enzyme can serve as a research tool for studying the metabolism of valacyclovir and structurally related compounds in controlled in vitro systems. Researchers might investigate the hydrolysis patterns of various prodrugs and examine how different chemical modifications affect substrate recognition and processing. Studies like these contribute to understanding human drug metabolism pathways and could inform medicinal chemistry research on prodrug design strategies.
