Recombinant Escherichia coli 1-acyl-sn-glycerol-3-phosphate acyltransferase (plsC) is produced using a yeast expression system and represents the full-length protein, spanning amino acids 1-245. The protein includes an N-terminal 6xHis-tag, which helps with purification and detection. SDS-PAGE analysis confirms a purity level greater than 90%, suggesting good reliability for research applications. This product is intended for research use only and is not suitable for therapeutic applications.
1-acyl-sn-glycerol-3-phosphate acyltransferase (plsC) appears to play a critical role in bacterial lipid biosynthesis, particularly in phospholipid formation. The enzyme catalyzes the acylation of lysophosphatidic acid to form phosphatidic acid, an essential precursor in membrane phospholipid biosynthesis. For researchers focusing on bacterial lipid metabolism and membrane dynamics, studying this enzyme may be crucial for understanding these complex 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.
1. Biochemical Characterization of Bacterial Phospholipid Biosynthesis
This recombinant plsC protein can be used to study the enzymatic properties and kinetic parameters of 1-acyl-sn-glycerol-3-phosphate acyltransferase in vitro. Researchers might investigate substrate specificity, optimal reaction conditions, and cofactor requirements for this key enzyme. The purified protein allows for detailed mechanistic studies of the second step in phosphatidic acid synthesis, which appears essential for bacterial membrane formation. Meanwhile, the N-terminal 6xHis tag makes protein purification and immobilization for enzymatic assays more straightforward.
2. Inhibitor Screening and Drug Discovery Research
The recombinant plsC protein may serve as a valuable target for screening potential antimicrobial compounds that could disrupt bacterial membrane biosynthesis. Since phospholipid synthesis seems essential for bacterial survival and this pathway differs from mammalian lipid metabolism, plsC likely represents an attractive target for antibacterial drug development research. High-throughput screening assays can be developed using this purified protein to identify small molecule inhibitors. The high purity level (>90%) should help ensure reliable and reproducible results in compound screening studies.
3. Protein-Protein Interaction Studies
Researchers can use this recombinant protein in pull-down assays and co-immunoprecipitation experiments to identify potential binding partners involved in bacterial lipid metabolism. The N-terminal 6xHis tag allows immobilization on nickel-affinity matrices for capturing interacting proteins from bacterial cell lysates. Such studies could reveal regulatory mechanisms and protein complexes involved in coordinating phospholipid biosynthesis with other cellular processes. These interaction studies may provide insights into the metabolic networks that appear to control bacterial membrane composition.
4. Structural Biology and Protein Folding Studies
The purified recombinant plsC protein can be used for structural characterization studies including X-ray crystallography, NMR spectroscopy, or cryo-electron microscopy. These approaches would likely provide detailed insights into the three-dimensional structure and catalytic mechanism of this important metabolic enzyme. The protein can also serve as a model system for investigating protein folding, stability, and conformational changes under different conditions. The yeast expression system may provide properly folded protein suitable for structural studies, though this would need experimental validation.
5. Antibody Development and Immunological Research
This recombinant protein can serve as an antigen for generating specific antibodies against E. coli plsC for research applications. The purified protein makes it possible to produce polyclonal or monoclonal antibodies that researchers can use in Western blotting, immunofluorescence, or immunohistochemistry studies of bacterial samples. Such antibodies would likely be valuable tools for studying plsC expression levels, subcellular localization, and regulation under different bacterial growth conditions. The N-terminal His tag can also be used in tag-specific detection methods during antibody validation.
