Recombinant Clostridium symbiosum Pyruvate, phosphate dikinase (ppdK) is produced in E. coli and includes both an N-terminal 10xHis tag and a C-terminal Myc tag for straightforward purification and detection. This partial protein covers amino acids 384 to 511 and is purified to over 90% homogeneity, as confirmed by SDS-PAGE. The product is designed solely for research use and appears to offer reliable quality and consistency for experimental applications.
Pyruvate, phosphate dikinase plays a critical role in metabolic pathways by catalyzing the conversion of pyruvate to phosphoenolpyruvate. This enzyme is integral to energy metabolism and carbon fixation processes, which makes it a crucial component of research into microbial metabolic pathways and enzymatic regulation. Understanding its function and mechanics seems vital for studies in bioenergetics and microbial physiology.
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. Antibody Development and Immunoassay Research
This recombinant ppdK fragment (384-511aa) with dual His and Myc tags can serve as an immunogen for generating specific antibodies against Clostridium symbiosum pyruvate, phosphate dikinase. The N-terminal His tag allows efficient purification for immunization protocols. Meanwhile, the C-terminal Myc tag provides an additional epitope for antibody validation studies. Researchers can use this protein to develop polyclonal or monoclonal antibodies for studying ppdK expression and localization in bacterial systems. The >90% purity likely makes it suitable for immunization without significant contaminating proteins that could generate cross-reactive antibodies.
2. Protein-Protein Interaction Studies
The dual-tagged nature of this recombinant protein makes it valuable for investigating potential protein partners of the ppdK C-terminal domain in bacterial metabolic pathways. The His tag allows for nickel-affinity pull-down experiments using bacterial lysates to identify interacting proteins. The Myc tag then confirms protein capture through Western blotting. This partial protein fragment may retain important protein-binding domains that could reveal novel regulatory mechanisms or metabolic complexes involving pyruvate, phosphate dikinase. High purity levels suggest minimal background binding in interaction assays.
3. Epitope Mapping and Structural Domain Analysis
This specific fragment (384-511aa) represents a defined region of the full-length ppdK protein. Scientists can use it to map functional or antigenic epitopes within this C-terminal domain. Researchers might perform limited proteolysis experiments, cross-linking studies, or hydrogen-deuterium exchange analysis to characterize the structural properties of this domain. The dual tags make detection and purification straightforward throughout these analytical procedures. Such studies could provide insights into the modular organization of pyruvate, phosphate dikinase and identify critical regions for enzyme function or regulation.
4. Comparative Biochemical Analysis
This recombinant fragment can serve as a reference standard for comparative studies of pyruvate, phosphate dikinase variants across different bacterial species or mutant strains. The standardized expression system and purification tags allow for consistent preparation and quantification of the protein for biochemical characterization experiments. Researchers can analyze thermal stability, pH sensitivity, or chemical modification patterns of this specific domain compared to corresponding regions from other organisms. The >90% purity appears to ensure reliable and reproducible results in comparative biochemical assays.
