Recombinant Hypoderma lineatum Hypodermin-A is produced in E. coli and contains the complete mature protein sequence spanning amino acids 31 to 256. The protein carries an N-terminal 6xHis tag, which makes purification and detection more straightforward. SDS-PAGE analysis shows the product achieves purity levels above 85%, which appears sufficient for most research applications.
Hypodermin-A is a protease that comes from the common cattle grub, Hypoderma lineatum. This protein seems to play an important role in how the parasite completes its life cycle, likely helping it invade tissues and obtain nutrients. Understanding this protein may be significant for grasping how parasitic mechanisms work and could potentially lead to better control strategies. This makes it a useful research tool in parasitology studies.
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.
Hypodermin-A is a serine protease from a parasitic insect that requires precise disulfide bond formation and proper tertiary structure for its enzymatic activity. The E. coli expression system cannot perform the necessary post-translational modifications (particularly proper disulfide bonding) that are critical for this protein's native conformation. While the protein may be soluble, it is highly unlikely to achieve the correct folding needed for functional protease activity. The probability of correct folding with enzymatic function is low.
1. Antibody Development and Immunological Studies
This recombinant Hypodermin-A serves as an excellent immunogen for generating antibodies against linear epitopes of the protein. The full-length mature sequence ensures comprehensive epitope coverage. The high purity minimizes antibodies against contaminants. However, antibodies may not efficiently recognize conformational epitopes on the native, properly folded enzyme.
2. Biochemical Characterization
This is the essential first step to assess physical properties. Techniques like circular dichroism can analyze secondary structure, while thermal shift assays evaluate stability. These studies provide critical data about the protein itself, not the native protein.
3. Comparative Parasitology Research
This protein can be used for sequence-based comparisons and immunological cross-reactivity studies across parasite species. However, comparative functional studies would be invalid due to the protein's likely misfolded state.
Final Recommendation & Action Plan
This recombinant Hypodermin-A is primarily suitable for antibody development and biochemical characterization but fundamentally unsuitable for functional studies due to E. coli's inability to produce properly folded enzyme. The immediate priority is Application 2 (Biochemical Characterization) to assess the protein's physical properties. Application 1 (Antibody Development) can proceed confidently. Protein interaction studies and functional studies must be avoided due to the high probability of misfolding. For functional protease studies, alternative approaches using eukaryotic expression systems or native purification from the parasite are essential.
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