Recombinant Escherichia coli 30S ribosomal protein S4 (rpsD) represents a full-length protein expressed in E. coli, spanning amino acids 2 to 206. This protein carries an N-terminal 6xHis tag for easier purification and detection. SDS-PAGE analysis indicates the product achieves greater than 90% purity, which appears to make it suitable for various research applications.
The 30S ribosomal protein S4 is a crucial component of the small ribosomal subunit in Escherichia coli. It plays what seems to be a central role in ribosome assembly and function. S4 participates in the translation process, helping to ensure accurate decoding of mRNA into proteins. Its interactions with rRNA and other ribosomal proteins may be essential for maintaining ribosomal structural integrity, making it an important focus in molecular biology and genetics 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. Ribosome Assembly and Biogenesis Studies
This recombinant S4 protein offers a way to investigate the sequential assembly pathway of the 30S ribosomal subunit in vitro. Researchers might examine how S4 interacts with 16S rRNA and other ribosomal proteins during early ribosome assembly stages. The N-terminal His-tag makes purification straightforward and allows tracking of S4 incorporation into ribosomal complexes through pull-down assays or immunodetection methods.
2. Protein-RNA Interaction Analysis
The recombinant S4 protein appears well-suited for studying its specific binding interactions with 16S rRNA, particularly at the 5' domain where S4 is known to bind. Electrophoretic mobility shift assays (EMSA) or surface plasmon resonance experiments could help characterize binding kinetics and specificity. The His-tag makes purification and immobilization for various binding assays more manageable.
3. Structural and Biochemical Characterization
This purified S4 protein might prove useful for detailed structural studies. X-ray crystallography, NMR spectroscopy, or cryo-electron microscopy could reveal its three-dimensional structure and conformational dynamics. The high purity (>90%) likely makes it suitable for biophysical analyses such as circular dichroism spectroscopy to study protein folding and stability under different conditions.
4. Antibody Development and Validation
The recombinant S4 protein could serve as an antigen for generating specific antibodies against E. coli ribosomal protein S4. These antibodies would likely prove valuable for immunoblotting, immunofluorescence, and immunoprecipitation experiments studying ribosome localization and dynamics in bacterial cells. The His-tag also enables ELISA-based assays for antibody screening and characterization.
5. Ribosome Reconstitution Experiments
This recombinant S4 protein can be incorporated into in vitro ribosome reconstitution systems to study minimal requirements for functional 30S subunit assembly. Researchers may systematically add or omit specific ribosomal components, including this S4 protein, to determine their individual contributions to ribosome function and assembly efficiency. Such studies could provide insights into the essential versus accessory roles of ribosomal proteins in translation machinery.
