Recombinant Aspergillus restrictus Ribonuclease mitogillin (ret) is produced in E. coli and contains the full-length mature protein from amino acids 28 to 176. The protein includes an N-terminal 10xHis-tag and a C-terminal Myc-tag, which makes purification and detection more straightforward. SDS-PAGE analysis indicates the protein's purity exceeds 85%, suggesting it may be suitable for reliable research applications. This product is intended for research use only.
Ribonuclease mitogillin from Aspergillus restrictus appears to be a ribonuclease enzyme recognized for its role in cleaving RNA molecules. Researchers commonly study it for its enzymatic activity and potential applications in molecular biology research. Understanding how such ribonucleases work and what they do could provide insights into RNA processing and degradation—processes that seem crucial in various cellular and molecular 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.
Ribonuclease mitogillin is a fungal ribotoxin that requires precise disulfide bond formation and specific tertiary structure for its enzymatic activity and target recognition. The E. coli expression system cannot provide the eukaryotic folding environment or post-translational modifications necessary for this complex protein. Additionally, the dual N-terminal and C-terminal tags may sterically interfere with proper folding and active site formation. While the protein may be soluble, the probability of correct folding with functional ribonuclease activity is extremely low.
1. Antibody Development and Immunoassay Optimization
This recombinant protein serves as an excellent immunogen for generating antibodies against linear epitopes of mitogillin. The dual tags provide additional epitopes for antibody screening. However, antibodies may not recognize conformational epitopes on the native, properly folded protein.
2. Comparative Structural and Biochemical Analysis
This is the essential first step to assess the protein's physical properties. Techniques like circular dichroism can analyze secondary structure, while size-exclusion chromatography can determine oligomeric state. These studies provide critical quality control data but characterize the protein itself, not the native protein.
Final Recommendation & Action Plan
The E. coli expression system is fundamentally unsuitable for producing a functional fungal ribonuclease, limiting applications to non-functional uses. The immediate priority is Application 2 (Biochemical Characterization) to assess the protein's physical state. Application 1 (Antibody Development) can proceed confidently for generating linear epitope antibodies. Ribonuclease interactions require precise tertiary structure. A misfolded, tagged variant will yield misleading results. Cellular localization is dependent on native structure and function, which this misfolded protein cannot provide. For functional studies, ribonuclease mitogillin from fungal expression systems or native purification is essential.
