Recombinant Human Receptor-type tyrosine-protein phosphatase delta (PTPRD) comes from a mammalian cell expression system and spans amino acid region 21-1265. The protein appears to be partially expressed and includes a C-terminal 10xHis-tag, which should make purification and detection more straightforward. Purity levels exceed 95% based on SDS-PAGE analysis, while endotoxin levels stay below 1.0 EU/ug. The protein shows biological activity and successfully binds Human IL1RAPL1 in functional ELISA assays.
Receptor-type tyrosine-protein phosphatase delta (PTPRD) participates in essential cellular processes, working mainly to dephosphorylate target proteins. This enzyme plays what seems to be a critical role in signaling pathways that control cell growth, differentiation, and neural development. PTPRD's catalytic activity and protein interactions have made it an important research target, particularly for scientists studying cellular communication and signal transduction mechanisms.
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 with IL1RAPL1
This application is well-supported and valid. The PTPRD protein has demonstrated specific, high-affinity binding to IL1RAPL1 with a characterized EC??. The C-terminal His-tag facilitates immobilization for detailed interaction studies using techniques like surface plasmon resonance (SPR) or bio-layer interferometry (BLI) to determine kinetic parameters (kon, koff, KD). This recombinant PTPRD protein is an excellent reagent for quantitatively characterizing the molecular mechanism of the PTPRD-IL1RAPL1 interaction in neuronal contexts.
2. Competitive Binding Assays for Interaction Mapping
This application is highly feasible. The validated ELISA binding assay provides a robust platform for competitive inhibition studies. The known EC?? serves as a precise baseline for screening small molecules or other proteins that compete with IL1RAPL1 for binding to PTPRD. This approach is ideal for identifying potential modulators of this specific interaction or for mapping whether other binding partners share the same or an overlapping binding site on PTPRD.
3. Antibody Development and Validation
This application is suitable. The high purity, low endotoxin levels, and confirmed native-like folding (inferred from its bioactivity) make this protein an ideal immunogen for generating high-quality antibodies. Crucially, because the protein is functional, antibodies can be screened not just for binding (e.g., by ELISA or Western blot) but also for their ability to block the PTPRD-IL1RAPL1 interaction in a functional assay, allowing for the development of function-blocking antibodies.
4. Biochemical Characterization of Phosphatase Activity
This application requires a critical caveat. The recombionant PTPRD protein's activity has been validated only for its binding function to IL1RAPL1. There is no data presented confirming its enzymatic (phosphatase) activity. While the construct includes the catalytic domain, its enzymatic function must be experimentally verified before any kinetic or substrate specificity studies can be meaningfully pursued. This protein could be used to characterize phosphatase activity, but this is contingent upon first confirming enzymatic function with a phosphatase activity assay.
5. Structure-Function Relationship Studies
This application is valid for binding function studies. The PTPRD protein is highly suitable for investigating the structural determinants of IL1RAPL1 binding (e.g., using domain mapping or limited proteolysis). Without confirmed phosphatase activity, structure-function conclusions cannot be drawn regarding the catalytic mechanism or its regulation. The focus should initially be on structure-binding relationships.
Final Recommendation & Action Plan
The immediate and highly recommended course of action is to leverage this well-characterized protein for the applications it has been validated for: detailed biophysical analysis of the IL1RAPL1 interaction (Application 1), competitive screening assays (Application 2), and antibody development (Application 3). For Application 5 (structure-function), initial studies should focus exclusively on mapping the IL1RAPL1 binding domain. Before attempting Application 4 (phosphatase characterization), it is essential to first conduct a phosphatase activity assay using a standard chromogenic or fluorogenic phosphotyrosine substrate to confirm that the recombinant protein is indeed enzymatically active. The high purity and confirmed folding for binding are positive indicators, but enzymatic activity is distinct and must be verified separately. Once phosphatase activity is confirmed, the protein becomes an exceptionally powerful tool for comprehensive mechanistic studies integrating both its receptor-binding and enzymatic functions.
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