Recombinant Epstein-Barr virus Glycoprotein 42 (BZLF2) is expressed in E. coli, spanning amino acids 34 to 223 of the viral protein. This partial protein carries a C-terminal 6xHis tag and reaches a purity level of over 90% as confirmed by SDS-PAGE analysis. The protein is designed for research use only, though it appears to offer a dependable option for researchers investigating viral proteins and their interactions.
Glycoprotein 42 (gp42) of the Epstein-Barr virus seems to play a crucial role in the virus's ability to infect host cells. The protein is involved in the fusion process, helping the virus enter the host by interacting with cellular receptors. Understanding gp42's function may be essential for studying viral entry mechanisms and pathogenesis, which likely makes it a significant focus in virology research.
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.
The recombinant EBV glycoprotein 42 (a viral glycoprotein) is expressed in E. coli, a prokaryotic system that lacks the machinery for eukaryotic-type post-translational modifications, such as glycosylation. Glycoprotein 42 is naturally glycosylated in viral contexts, and glycosylation is often critical for correct folding, stability, and biological activity (e.g., receptor binding). The protein is expressed as a partial fragment (34-223aa), which may not include full structural domains required for native conformation. Since activity is unverified, the protein is likely misfolded or inactive due to the absence of glycosylation and potential improper folding in E. coli. Therefore, it cannot be assumed to be correctly folded or bioactive without experimental validation.
1. Antibody Development and Characterization
This recombinant EBV glycoprotein 42 fragment can work as an immunogen for generating antibodies, but if misfolded (which is probable due to lack of glycosylation), the antibodies may primarily recognize linear epitopes and might not bind the native, glycosylated form of the protein in viral contexts. The His tag aids purification, and >90% purity is sufficient for immunization. However, antibodies should be validated against native glycoprotein 42 from EBV-infected cells to ensure relevance to physiological conditions.
2. Protein-Protein Interaction Studies
The use of this recombinant glycoprotein 42 in pull-down assays is high-risk. If the protein is misfolded (as expected due to expression in E. coli), it may not interact authentically with host receptors or viral partners, leading to false positives or negatives. The His tag allows immobilization, but the results would be unreliable for studying pathogenesis without folding validation. This application should be avoided unless correct folding is confirmed.
3. ELISA-Based Binding Assays
This application is problematic if the assay depends on conformational epitopes. The recombinant protein can be coated on plates, but if misfolded, binding studies with receptors (e.g., MHC class II) may not reflect native interactions. The high purity improves coating efficiency, but data should be interpreted cautiously. If used for linear epitope mapping, it might work, but for conformational studies, it is not recommended without folding checks.
4. Western Blot Standard and Positive Control
This application is suitable because Western blotting operates under denaturing conditions, where protein folding is irrelevant. The recombinant protein can serve as a positive control for size reference and antibody validation against the linear sequence. The partial fragment (34-223aa) and His tag provide a defined standard, and high purity ensures reliability.
Final Recommendation & Action Plan
Given the high likelihood of misfolding due to E. coli expression lacking glycosylation, it is essential to validate protein folding before any bioactive applications. Recommend first performing biophysical analyses (e.g., circular dichroism for secondary structure, size-exclusion chromatography for oligomeric state) to assess folding. If misfolding is confirmed, prioritize applications like antibody development (for linear epitopes) and Western blot standards, while avoiding interaction or binding studies. For functional insights, consider alternative expression systems (e.g., mammalian cells) that support glycosylation. Always include controls comparing with the native protein when possible.
