Recombinant Human Glycogen Phosphorylase, brain form (PYGB), is produced in E. coli and includes the 6-843 amino acid region. This partial protein carries an N-terminal GST tag and shows purity exceeding 90% when measured by SDS-PAGE. The product is intended for research use only, offering what appears to be a dependable tool for studies that require high-purity recombinant proteins.
Glycogen phosphorylase, brain form (PYGB), plays a crucial role in glycogen metabolism by catalyzing the breakdown of glycogen to glucose-1-phosphate. This process seems essential for maintaining energy homeostasis in brain tissue, particularly under conditions of high energy demand. PYGB's involvement in energy metabolism makes it an important focus of research in neurobiology and related metabolic 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.
1. Glycogen Metabolism Enzyme Activity Assays
This recombinant PYGB protein can serve as a positive control or reference standard in biochemical assays studying glycogen phosphorylase activity and glycogen metabolism pathways. The N-terminal GST tag helps with protein purification and immobilization for enzyme kinetic studies. Researchers might use this protein to establish baseline activity measurements and optimize assay conditions for glycogen breakdown studies. The high purity (>90%) likely makes it suitable for quantitative biochemical analyses where protein concentration accuracy is critical.
2. Antibody Development and Validation
The GST-tagged recombinant PYGB can be used as an immunogen for generating specific antibodies against human glycogen phosphorylase brain form. The protein may serve as a positive control in Western blot, ELISA, and immunoprecipitation experiments to validate antibody specificity and sensitivity. The GST tag allows for straightforward purification and immobilization on glutathione-based matrices for antibody screening and characterization. This application appears particularly valuable for developing research tools to study PYGB expression and localization in various cell types and tissues.
3. Protein-Protein Interaction Studies
The GST-tagged PYGB protein can be immobilized on glutathione beads for pull-down assays to identify potential binding partners and regulatory proteins that interact with glycogen phosphorylase. The tag simplifies capture and isolation of protein complexes from cell lysates or purified protein mixtures. This approach might help clarify the molecular mechanisms governing glycogen phosphorylase regulation and its role in metabolic networks. The high purity of the protein should minimize background interactions in these binding studies.
4. Structural and Biophysical Characterization
This recombinant PYGB protein can be used in structural biology studies including X-ray crystallography, NMR spectroscopy, or cryo-electron microscopy to understand the three-dimensional structure of human brain glycogen phosphorylase. The protein appears suitable for biophysical analyses such as dynamic light scattering, analytical ultracentrifugation, and thermal stability studies. The GST tag can be removed by protease cleavage if needed for structural studies, or the tagged version can be used to study the effects of N-terminal modifications on protein folding and stability.
5. Drug Discovery and Compound Screening
The recombinant PYGB protein may serve as a target for screening small molecule libraries to identify potential modulators of glycogen phosphorylase activity in research settings. The GST tag enables development of homogeneous assay formats for high-throughput screening applications. Researchers can use this protein to study structure-activity relationships of known glycogen phosphorylase inhibitors or activators. The high purity should ensure reliable and reproducible results in dose-response studies and mechanism of action investigations.
