Recombinant Staphylococcus aureus 30 kDa neutral phosphatase gets produced in E.coli and comes with an N-terminal 6xHis-SUMO tag that helps with purification and appears to boost stability. This partial protein covers the 1-35 amino acid region and reaches over 90% purity, which SDS-PAGE analysis confirms. It's designed strictly for research purposes and seems to offer a dependable, high-quality choice for studies that need this particular phosphatase.
The 30 kDa neutral phosphatase from Staphylococcus aureus is an enzyme that likely plays a key role in phosphate metabolism. Phosphatases are important for many cellular processes - signal transduction and energy metabolism among them - by removing phosphate groups from substrates. Studying this enzyme may provide insights into bacterial physiology and potential interactions within host environments. This makes it a potentially valuable tool in microbiological and biochemical research.
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. Protein-Protein Interaction Studies Using Pull-Down Assays
The N-terminal 6xHis-SUMO tag allows researchers to immobilize this recombinant S. aureus phosphatase fragment on nickel-affinity resins for pull-down experiments. Scientists can then investigate potential binding partners or interacting proteins from S. aureus cell lysates or other bacterial extracts. The partial protein sequence (amino acids 1-35) might contain important binding domains or regulatory regions that interact with other cellular components. This approach could help reveal the role of the N-terminal region in protein complex formation or cellular localization.
2. Antibody Development and Epitope Mapping
The purified recombinant protein fragment can work as an immunogen for generating polyclonal or monoclonal antibodies specific to the N-terminal region of S. aureus 30 kDa neutral phosphatase. The high purity (>90%) appears to minimize contamination that could lead to cross-reactive antibodies. These antibodies could then be used in follow-up research applications like Western blotting, immunofluorescence, or immunoprecipitation studies. The partial protein represents a defined antigenic region that may be less accessible in the full-length native protein.
3. Structural and Biochemical Characterization Studies
Researchers can analyze the N-terminal fragment using various biophysical techniques to understand the structural properties of this specific region of the phosphatase. Techniques like circular dichroism spectroscopy, dynamic light scattering, or NMR could provide insights into the secondary structure and folding characteristics of amino acids 1-35. The SUMO tag can be removed by specific proteases if structural studies require it. These analyses might contribute to understanding the overall architecture and domain organization of the full-length enzyme.
4. ELISA-Based Binding Assays
The His-SUMO tagged protein fragment can be used to develop enzyme-linked immunosorbent assays for studying binding interactions with potential ligands, substrates, or regulatory molecules. The protein can be immobilized directly on ELISA plates or captured using anti-His or anti-SUMO antibodies for oriented immobilization. This system allows for quantitative measurement of binding affinities and kinetic parameters. Such assays could prove useful for screening small molecule libraries or investigating the binding properties of the N-terminal domain.
