Recombinant Mouse Procollagen-lysine,2-oxoglutarate 5-dioxygenase 2 (Plod2) gets expressed in a baculovirus system. The protein covers the full-length mature sequence from amino acids 26-737. A C-terminal 6xHis-Myc tag is attached to make purification and detection more straightforward. SDS-PAGE analysis shows the purity exceeds 85%, which appears to provide reliable performance for research work.
Plod2 functions as an enzyme that's crucial for collagen biosynthesis. Its main job involves hydroxylating lysine residues in collagen-like peptides. This modification seems essential for maintaining collagen fiber stability and function, which directly affects connective tissue integrity. Research interest in Plod2 likely stems from its role in extracellular matrix formation and maintenance pathways.
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
Researchers can use this recombinant Plod2 protein to study the enzymatic properties of procollagen-lysine,2-oxoglutarate 5-dioxygenase 2 under controlled laboratory conditions. Scientists might investigate kinetic parameters like Km and Vmax values by testing different procollagen substrates along with cofactors such as 2-oxoglutarate, ascorbate, and iron. The C-terminal His-Myc tag makes protein purification and quantification easier, helping determine accurate enzyme concentrations for kinetic experiments. These studies could provide fundamental biochemical data about mouse Plod2's catalytic efficiency and which substrates it prefers.
2. Protein-Protein Interaction Mapping
The dual C-terminal His-Myc tag system opens up possibilities for comprehensive protein interaction studies through both affinity purification and immunoprecipitation methods. Scientists can run pull-down assays using the His tag to discover new binding partners of Plod2 from mouse cell lysates or tissue extracts. Meanwhile, the Myc tag works well for immunoprecipitation experiments and Western blot detection to confirm specific protein interactions. This strategy may reveal regulatory proteins or cofactors that influence how Plod2 functions in collagen biosynthesis pathways.
3. Antibody Development and Validation
This purified recombinant protein works as an ideal antigen for creating specific antibodies against mouse Plod2. The high purity level (>85%) suggests minimal cross-reactivity with other proteins during immunization protocols. Scientists can use this protein as both a positive control and standard when validating antibody specificity in Western blotting, immunofluorescence, and immunohistochemistry work. The known molecular weight and tag information help with proper antibody characterization and optimizing detection conditions.
4. Structural and Biophysical Characterization
The full-length mature protein (26-737aa) from the baculovirus expression system may provide a suitable sample for structural biology studies. These could include X-ray crystallography, NMR spectroscopy, or cryo-electron microscopy. Scientists might investigate mouse Plod2's three-dimensional structure and compare it with related family members to better understand structure-function relationships. The protein also appears useful for biophysical studies like dynamic light scattering, thermal stability assays, and circular dichroism spectroscopy to characterize folding properties and stability under different conditions.
5. Inhibitor Screening and Drug Discovery Research
This recombinant Plod2 protein could serve as a target for high-throughput screening of potential enzyme inhibitors in drug discovery research focused on collagen-related disorders. Scientists can develop biochemical assays to test compound libraries for inhibitory activity against Plod2's enzymatic function. The tagged protein allows for easier detection and quantification in plate-based assay formats, which may help identify promising lead compounds. Such studies would likely contribute to understanding Plod2 as a potential therapeutic target and provide tools for additional preclinical research.
