Recombinant Mouse Leucine-rich repeat and immunoglobulin-like domain-containing nogo receptor-interacting protein 1 (Lingo1) is produced in a yeast expression system, covering the extracellular domain from amino acids 37 to 555. The protein includes an N-terminal 6xHis tag for purification and detection. SDS-PAGE analysis confirms purity greater than 90%, which appears to make it suitable for various experimental applications in research settings.
Lingo1 seems to play a critical role in neuronal development and regeneration. This protein is involved in pathways that regulate axon guidance and myelination. It acts as a key inhibitor of axonal growth. Researchers studying neurobiological processes find Lingo1 particularly interesting, especially when investigating conditions related to nerve injury and degeneration.
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.
Based on the provided information, the folding state and bioactivity of this recombinant mouse Lingo1 protein are unknown and cannot be assumed. Lingo1 is a transmembrane protein containing leucine-rich repeats (LRR) and immunoglobulin-like domains, which require precise folding and disulfide bond formation for proper function. Expression in a yeast system (eukaryotic) is favorable for disulfide bond formation compared to prokaryotic systems, but the presence of an N-terminal 6xHis tag may potentially interfere with the N-terminal structure or folding. The extracellular domain (37-555aa) is expressed, but without activity validation, it is uncertain whether the protein adopts a native-like conformation capable of binding to its known partners (e.g., Nogo receptor or other ligands). The >90% purity indicates minimal contaminants but does not confirm correct tertiary structure or bioactivity. Therefore, applications relying on specific biological interactions are speculative without experimental validation.
1. Protein-Protein Interaction Studies Using His-Tag Pull-Down Assays
The N-terminal 6xHis tag allows for immobilization on nickel-affinity resins for pull-down experiments. However, the utility for identifying biological interaction partners is entirely contingent on the recombinant Lingo1 protein being correctly folded. The LRR and Ig domains are conformation-dependent; if misfolded, they may not present native binding surfaces, leading to non-specific interactions or false negatives. Results from such assays should be considered preliminary and require validation with a bioactive protein or orthogonal methods.
2. Antibody Development and Validation
This recombinant Lingo1 extracellular domain is suitable as an immunogen for generating antibodies. The yeast expression system may promote proper folding, increasing the likelihood of antibodies recognizing conformational epitopes. However, without activity validation, it is uncertain whether the protein is natively folded. Antibodies generated may primarily recognize linear epitopes or the His-tag, and their ability to bind the full-length, membrane-associated Lingo1 in mouse tissues must be empirically validated. The protein can be used as a positive control in immunoassays, but results should be interpreted with caution.
3. Biochemical Characterization and Biophysical Analysis
This purified recombinant Lingo1 protein is well-suited for biochemical and biophysical characterization, including size exclusion chromatography, dynamic light scattering, thermal stability assays, and circular dichroism spectroscopy. This application is valid as it focuses on intrinsic physical properties (e.g., oligomerization state, stability) that are independent of bioactivity. However, the data characterize this specific recombinant Lingo1 protein, and findings may not fully represent the native Lingo1 protein due to the His-tag and expression system.
4. Cell-Based Binding and Localization Studies
This application is not recommended without prior validation of bioactivity. Using this recombinant Lingo1 protein in cell-based binding studies presupposes correct folding and functionality. If the protein is misfolded, any observed binding or localization may be artifactual and not reflect physiological interactions. Fluorescent labeling or His-tag detection could be used, but results would be uninterpretable in a biological context without confirmation of native structure.
Final Recommendation & Action Plan
The immediate priority is to experimentally validate the folding and bioactivity of this recombinant Lingo1 protein before proceeding with functional studies. This can be done using techniques such as circular dichroism to assess secondary structure, size-exclusion chromatography to check for proper oligomerization, and functional assays (e.g., binding to known partners like the Nogo receptor using surface plasmon resonance or cell-based assays). If bioactivity is confirmed, the protein becomes valuable for interaction studies (Application 1), antibody development (Application 2), and cell-based studies (Application 4). If inactive, its use should be restricted to biochemical characterization (Application 3) and as an immunogen for generating linear-epitope antibodies (Application 2, with limitations). Until validated, applications relying on native conformation should be considered exploratory, and results interpreted with caution.
