Recombinant Rat Pyruvate kinase PKM is produced in a baculovirus expression system, covering the full-length protein sequence from amino acids 1 to 531. It features an N-terminal 10xHis-tag and a C-terminal Myc-tag, ensuring efficient purification and detection. The protein is purified to over 85% purity, as verified by SDS-PAGE, and is designed for research use only. This appears to provide a reliable tool for experimental applications.
Pyruvate kinase PKM plays a crucial role in glycolysis, catalyzing the conversion of phosphoenolpyruvate to pyruvate with the generation of ATP. This enzyme is essential for energy production and participates in various metabolic pathways. Its activity and regulation seem critical for understanding metabolic processes, which may make it a valuable target in research focused on cellular energy dynamics and metabolic diseases.
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 Metabolic Pathway Studies
This recombinant rat pyruvate kinase can help investigate glycolytic enzyme kinetics and metabolic flux analysis in controlled in vitro systems. Researchers might examine substrate specificity, cofactor requirements, and allosteric regulation mechanisms by measuring the conversion of phosphoenolpyruvate and ADP to pyruvate and ATP under various experimental conditions. The full-length protein (1-531aa) expressed in baculovirus system likely provides a eukaryotic post-translational modification profile that closely mimics native rat PKM. Comparative studies between different PKM isoforms or mutant variants could potentially elucidate structure-function relationships in glycolytic regulation.
2. Protein-Protein Interaction Screening
The dual-tagged design with N-terminal His-tag and C-terminal Myc-tag makes this protein suitable for pull-down assays and co-immunoprecipitation experiments to identify PKM binding partners. The His-tag allows immobilization on nickel-based resins for affinity purification of interacting proteins from rat tissue lysates or cell extracts. Meanwhile, the Myc-tag provides an additional detection and capture method for validation experiments and Western blot analysis. These interaction studies may reveal novel regulatory networks involving pyruvate kinase in cellular metabolism.
3. Antibody Development and Validation
This recombinant protein appears to serve as an excellent immunogen and standard for developing rat PKM-specific antibodies for research applications. The high purity (>85%) and full-length sequence likely ensure proper epitope presentation for generating polyclonal or monoclonal antibodies with high specificity. The protein can be used as a positive control in Western blotting, immunofluorescence, and ELISA-based detection systems. Researchers might validate antibody cross-reactivity and establish quantitative detection methods for PKM in rat tissue samples and cell culture studies.
4. Structural and Biophysical Characterization
The recombinant rat PKM can be applied in structural biology studies including X-ray crystallography, NMR spectroscopy, and cryo-electron microscopy to understand conformational changes and allosteric mechanisms. Biophysical techniques such as dynamic light scattering, differential scanning calorimetry, and surface plasmon resonance may provide insights into protein stability, oligomerization states, and ligand binding kinetics. The baculovirus expression system appears to ensure proper protein folding and post-translational modifications necessary for native-like structural studies.
5. Comparative Species Analysis and Evolution Studies
This rat PKM protein enables comparative biochemical studies across different mammalian species to understand evolutionary conservation and divergence of glycolytic enzymes. Researchers can perform side-by-side kinetic analyses with human, mouse, or other mammalian PKM orthologs to identify species-specific regulatory mechanisms. The protein may serve as a reference standard in phylogenetic studies examining metabolic enzyme evolution and adaptation in different mammalian lineages.
