Recombinant Rat Protein-lysine 6-oxidase (Lox) is produced in E. coli and spans the full length of the mature protein from amino acids 163 to 411. This high-purity preparation exceeds 90% purity as confirmed by SDS-PAGE and includes an N-terminal 6xHis-tag for easier purification and detection. The recombinant protein is designed strictly for research applications and appears suitable for various experimental approaches.
Protein-lysine 6-oxidase (Lox) serves as a key enzyme for cross-linking collagen and elastin, which means it plays a significant role in maintaining structural integrity of the extracellular matrix. The enzyme's activity seems essential during connective tissue remodeling and has become a central focus in studies examining tissue development and repair. Many researchers explore Lox within the context of fibrosis and cancer metastasis, given its apparent involvement in these critical biological 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. Protein-Protein Interaction Studies Using Pull-Down Assays
The N-terminal 6xHis-tagged recombinant rat Lox protein can be immobilized on nickel-affinity resins to identify potential binding partners from rat tissue lysates or cell extracts. This strategy may help clarify the molecular interactions of Lox in extracellular matrix formation and collagen processing pathways. With its high purity (>90%), there should be minimal background binding from contaminating proteins during pull-down experiments. Mass spectrometry analysis of co-precipitated proteins could potentially reveal novel Lox-interacting proteins in rat models.
2. Antibody Development and Validation
This recombinant rat Lox protein appears to be an ideal antigen for generating specific antibodies against the mature form of the enzyme. The defined expression region (163-411aa) represents the full-length mature protein, which suggests that resulting antibodies will likely recognize the biologically relevant form of Lox. Both the high purity and His-tag allow for straightforward purification and quantification needed for immunization protocols. Generated antibodies can then be validated using ELISA, Western blot, and immunoprecipitation assays with this same recombinant protein serving as a positive control.
3. Biochemical Characterization and Enzyme Kinetics Studies
The recombinant protein may prove useful for investigating the biochemical properties of rat Lox, including optimal pH, temperature stability, and cofactor requirements. Researchers can perform substrate specificity studies using various lysine and hydroxylysine-containing peptides or proteins to better understand the enzyme's catalytic preferences. The purified protein allows for controlled in vitro experiments to determine kinetic parameters and inhibitor sensitivity profiles. Such studies would likely provide fundamental biochemical data for rat Lox that can be compared across different species.
4. Structural Biology Applications
The high-purity recombinant rat Lox protein can be applied to structural studies including X-ray crystallography, NMR spectroscopy, or cryo-electron microscopy. The His-tag makes possible additional purification steps that may be necessary for structural biology work. Comparative structural analysis with Lox from other species could reveal evolutionary conservation and species-specific differences in active site architecture. These structural insights would likely enhance our understanding of the catalytic mechanism and substrate recognition patterns of mammalian lysyl oxidases.
