Recombinant Absidia glauca Actin-1 (ACT1) is produced in E. coli and expressed as a partial length protein (1-140 amino acids). The protein carries an N-terminal 6xHis-tag and a C-terminal Myc-tag, which streamline purification and detection processes. SDS-PAGE analysis confirms that the protein achieves a purity level exceeding 85%, suggesting reliable performance in research applications.
Actin-1 from Absidia glauca appears to be a key component of the cytoskeleton, playing what seems to be a crucial role in cell structure and motility. Actin proteins are integral to various cellular processes. These include maintaining cell shape, enabling intracellular transport, and supporting cell division. Such functions make actin an important focus in studies related to cell biology and dynamics.
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. Comparative Actin Structure-Function Studies
This recombinant Absidia glauca ACT1 fragment (1-140aa) may serve as a valuable tool for comparative studies examining actin evolution and structural diversity across different organisms. The N-terminal region of actin contains critical domains involved in nucleotide binding and polymerization. This makes the fragment suitable for investigating species-specific differences in actin biochemistry, though the extent of these differences remains to be fully characterized. Researchers can use this protein to compare binding affinities with various actin-binding proteins from different species. The dual His and Myc tags should simplify purification and detection in comparative binding assays.
2. Antifungal Drug Target Validation
The recombinant ACT1 fragment can be used in biochemical screens to identify compounds that specifically interact with fungal actin versus mammalian actin. This application leverages what appear to be evolutionary differences between fungal and animal actins to discover selective antifungal agents. The protein can be immobilized using the His-tag for high-throughput compound screening assays. Meanwhile, the Myc tag allows detection and quantification of protein-compound interactions through immunoassays.
3. Antibody Development and Characterization
This recombinant protein appears to serve as an excellent immunogen and screening antigen for developing antibodies specific to Absidia glauca or related fungal species. The dual tagging system allows for both purification via His-tag affinity chromatography and subsequent detection using anti-Myc antibodies in various immunoassays. Researchers can use this protein in ELISA-based screening to identify antibodies with desired specificity profiles. The fragment represents a defined antigenic region that may contain species-specific epitopes useful for diagnostic antibody development, though this will need experimental validation.
4. Protein-Protein Interaction Studies
The tagged ACT1 fragment can be used in pull-down assays to identify novel binding partners specific to fungal actin. The His-tag allows immobilization on nickel-based resins for affinity purification experiments using fungal cell lysates or purified protein libraries. The Myc tag provides an additional detection method for confirming protein interactions through co-immunoprecipitation studies. This approach might reveal unique protein interaction networks in pin mould that differ from well-characterized mammalian actin interactions.
5. Biochemical Characterization of Fungal Actin Properties
This recombinant fragment can be used to investigate the intrinsic biochemical properties of Absidia glauca actin, including nucleotide binding characteristics and stability under various conditions. The protein can be subjected to biophysical analyses such as circular dichroism spectroscopy to examine its folding properties and thermal stability. The dual tags should help with protein quantification and tracking during biochemical assays, allowing precise measurement of protein concentrations and recovery rates. These studies could provide fundamental insights into the molecular properties of fungal actins compared to their mammalian counterparts, though significant gaps in our understanding remain.
