Recombinant Neosartorya fumigata Vacuolar protease A (pep2) is expressed in E.coli, covering the mature protein region from amino acids 71 to 398. This product features an N-terminal 6xHis-tag, which makes purification and detection more straightforward. The protein achieves a purity level greater than 85% as confirmed by SDS-PAGE analysis. It is intended for research use only, without specified endotoxin levels or activity data.
Vacuolar protease A appears to be an important enzyme involved in protein processing and turnover within the fungal vacuole. It likely plays a critical role in the degradation of misfolded or damaged proteins, contributing to cellular homeostasis. Research on this protease may help scientists understand fungal biology better and could provide insights into proteolytic pathways relevant to broader biological systems.
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. Biochemical Characterization of Fungal Vacuolar Protease Activity
This recombinant N. fumigata vacuolar protease A can be used to establish in vitro protease activity assays using fluorogenic or chromogenic peptide substrates. Researchers might determine optimal pH conditions, temperature stability, and kinetic parameters to understand the enzyme's biochemical properties. The N-terminal 6xHis tag makes purification and immobilization easier for enzyme kinetics studies. Such characterization would likely provide fundamental insights into fungal vacuolar protease function and substrate specificity.
2. Antifungal Drug Target Validation Studies
The recombinant protein serves as a valuable tool for screening potential protease inhibitors as antifungal compounds in preclinical research. Scientists can perform high-throughput screening assays to identify small molecules that inhibit the protease activity. The purified protein allows for structure-activity relationship studies of inhibitor compounds and assessment of their specificity against fungal versus human proteases. This application may support the development of novel antifungal therapeutic strategies targeting essential fungal enzymes.
3. Antibody Development and Immunological Studies
The 6xHis-tagged recombinant protein can work as an immunogen for generating polyclonal or monoclonal antibodies specific to N. fumigata vacuolar protease A. These antibodies would be valuable research tools for studying protein localization, expression levels, and function in fungal cells. The purified protein also works as a positive control and standard in Western blotting, ELISA, and immunofluorescence experiments. Such immunological tools appear essential for investigating the role of this protease in fungal pathogenesis and cellular processes.
4. Protein-Protein Interaction Studies
The N-terminal 6xHis tag allows for pull-down assays to identify potential protein partners that interact with vacuolar protease A in fungal cellular processes. Researchers can use the immobilized recombinant protein to capture interacting proteins from fungal cell lysates, followed by mass spectrometry identification. This approach may help elucidate the protein's role in cellular pathways and its integration within the fungal proteolytic network. Understanding these interactions could provide insights into the enzyme's regulation and cellular function.
5. Comparative Protease Studies Across Fungal Species
This recombinant N. fumigata protease serves as a reference protein for comparative biochemical studies with homologous proteases from other fungal species. Scientists can analyze evolutionary relationships, substrate preferences, and functional differences between vacuolar proteases across different fungi. The standardized recombinant format allows for direct comparison of enzymatic properties and inhibitor sensitivities. Such comparative studies may contribute to understanding fungal protease evolution and species-specific adaptations.
