Recombinant Saccharomyces cerevisiae Sterol-4-alpha-carboxylate 3-dehydrogenase, decarboxylating (ERG26) is expressed in E. coli and contains the complete sequence spanning amino acids 1-349. The protein carries an N-terminal 10xHis-tag and a C-terminal Myc-tag, which streamline purification and detection processes. SDS-PAGE analysis indicates purity levels exceeding 90%, making it well-suited for rigorous research applications.
ERG26 appears to play a fundamental role in the ergosterol biosynthesis pathway—a process that seems critical for yeast and fungi survival. This enzyme catalyzes specific reactions in sterol intermediate conversion, ultimately contributing to ergosterol production. Since ergosterol is an essential membrane component in yeast cells, this protein may prove valuable for investigating sterol metabolism and could help researchers understand broader biological mechanisms in yeast 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 Sterol Biosynthesis Pathway
This recombinant ERG26 protein offers opportunities to examine the enzymatic properties and kinetic behavior of sterol-4-alpha-carboxylate 3-dehydrogenase under controlled laboratory conditions. Scientists can explore substrate preferences, cofactor dependencies, and optimal reaction environments using these purified enzyme preparations. The dual tagging system simplifies protein purification and detection, which should enable thorough biochemical analysis of this important enzyme in ergosterol production. Results from such investigations might enhance our understanding of yeast sterol metabolism and reveal the molecular mechanisms behind this dehydrogenase/decarboxylase function.
2. Protein-Protein Interaction Studies
Both the N-terminal His tag and C-terminal Myc tag make this protein well-suited for pull-down experiments aimed at discovering potential binding partners or regulatory proteins that may interact with ERG26. These tags allow for protein immobilization on suitable affinity matrices and detection within complex biological samples. Interaction studies like these could potentially uncover new regulatory pathways in sterol biosynthesis or identify proteins that might influence ERG26 activity. The high purity of this recombinant protein suggests it should produce dependable results in co-immunoprecipitation or similar interaction-based experiments.
3. Antibody Development and Validation
This highly purified recombinant ERG26 protein works well as an antigen for creating specific antibodies targeting the sterol-4-alpha-carboxylate 3-dehydrogenase enzyme. Since it contains the full-length protein sequence (1-349aa), researchers get comprehensive epitope coverage for developing both polyclonal and monoclonal antibodies. The dual-tag design makes purification straightforward and supports quality control throughout antibody development. Once generated, these antibodies could prove useful for Western blotting, immunofluorescence, or other detection techniques in yeast cell biology studies.
4. Structural and Biophysical Analysis
Researchers can use the recombinant ERG26 protein for structural investigations through X-ray crystallography, NMR spectroscopy, or cryo-electron microscopy to determine the three-dimensional architecture of this dehydrogenase enzyme. Various biophysical characterization methods—dynamic light scattering, circular dichroism spectroscopy, and thermal stability assays—may provide valuable information about protein folding patterns, stability characteristics, and conformational shifts. The high purity and dual-tag configuration should make protein handling and concentration measurements more manageable for these analytical techniques.
5. Enzyme Inhibitor Screening Assays
This purified ERG26 protein can serve as a target in high-throughput screening systems designed to identify compounds that might inhibit sterol-4-alpha-carboxylate 3-dehydrogenase activity. The recombinant enzyme becomes a testing ground for compound libraries in drug discovery research, particularly for developing antifungal agents. Both His and Myc tags allow for straightforward protein immobilization and detection across different assay formats. Screening efforts like these could potentially lead to discovering new chemical structures that disrupt yeast sterol biosynthesis for research applications.
