Recombinant Mouse Extracellular tyrosine-protein kinase PKDCC (Pkdcc) is expressed in a baculovirus system, spanning the full length of the mature protein from amino acids 33 to 492. The protein includes an N-terminal 10xHis-tag and a C-terminal Myc-tag, which makes purification and detection more straightforward. SDS-PAGE analysis shows purity exceeding 85%, suggesting it's well-suited for various research applications.
Extracellular tyrosine-protein kinase PKDCC appears to play a crucial role in cellular signaling pathways. The protein is known for its kinase activity and participates in phosphorylation processes that regulate various cellular functions. Studying this protein may be important for understanding signal transduction mechanisms and its potential involvement in cellular development and differentiation processes. Researchers use this protein to explore its functions and interactions within these 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.
Mouse PKDCC is an extracellular tyrosine kinase that requires precise folding, proper disulfide bond formation, and specific catalytic domain organization for its enzymatic activity. The baculovirus-insect cell expression system provides a eukaryotic environment that supports proper protein folding, disulfide bond formation, and some post-translational modifications. However, as an extracellular kinase, PKDCC may require specific glycosylation patterns and proper catalytic domain orientation that could be affected by the dual N-terminal 10xHis-tag and C-terminal Myc-tag. While the expression system increases folding probability, experimental validation is essential to confirm structural integrity and kinase activity.
1. Protein-Protein Interaction Studies Using Pull-Down Assays
This application carries a significant risk without proper folding validation. PKDCC interactions require precise tertiary structure and functional kinase domains. The dual tags may sterically interfere with binding interfaces or create artificial interaction surfaces. If correctly folded and active (verified), limited interaction studies may be possible. If misfolded/inactive (unverified), there is a high risk of non-specific binding or failure to identify genuine physiological interactions.
2. Antibody Development and Validation
This application is highly suitable regardless of folding status. Antibody development relies on antigenic sequence recognition rather than functional kinase activity. The full-length mature protein provides comprehensive epitope coverage for generating both linear and conformational antibodies. The high purity (>85%) ensures minimal contamination-related issues in immunization protocols.
3. Biochemical Characterization and Enzyme Kinetics Analysis
These studies are essential priority applications for determining folding status and functional competence. Comprehensive analysis should include size-exclusion chromatography to assess oligomeric state, circular dichroism spectroscopy to evaluate secondary structure, and kinase activity assays to validate catalytic function. However, the dual tags may interfere with substrate access or allosteric regulation in kinetic studies.
Final Recommendation & Action Plan
The baculovirus expression system provides favorable eukaryotic folding conditions for this extracellular kinase, but the dual-tag configuration and complex functional requirements necessitate rigorous validation before biological applications. Begin with Application 3 (Biochemical Characterization) to assess folding quality through SEC, CD spectroscopy, and validate kinase activity using appropriate substrates. Once correct folding and catalytic activity are verified, proceed cautiously with Application 1 for interaction studies. Application 2 (antibody development) can proceed immediately. Avoid cell-based assays entirely without functional validation, as misfolded kinases could yield misleading biological data. Always include appropriate controls: validate key findings with tag-free constructs when possible, use known substrates for activity confirmation, and consider the tags' potential interference in data interpretation. For critical functional studies, consider using a mammalian-expressed protein with minimal tags to ensure physiological relevance.
