Recombinant Human Isocitrate dehydrogenase [NAD] subunit gamma, mitochondrial (IDH3G) is produced in E. coli and represents the full-length mature protein from amino acids 40 to 393. The protein carries an N-terminal 6xHis-SUMO tag, which makes purification and detection more straightforward. SDS-PAGE analysis shows purity levels exceeding 90%, indicating the material should be suitable for research applications.
Isocitrate dehydrogenase [NAD] subunit gamma appears to be a crucial component of the mitochondrial enzyme complex that drives the citric acid cycle—a fundamental metabolic pathway. The protein helps catalyze the oxidative decarboxylation of isocitrate, which contributes to energy production through NADH generation. Given its role in cellular respiration, IDH3G has become a significant focus in metabolic 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
This recombinant IDH3G protein may be useful for investigating how it interacts with other subunits of the NAD-dependent isocitrate dehydrogenase complex (IDH3A and IDH3B). Researchers can take advantage of the N-terminal His tag for co-immunoprecipitation or pull-down assays. The purified protein should allow scientists to study how the heterotrimeric IDH3 complex assembles and identify regulatory proteins that might interact with the gamma subunit. In vitro binding assays could help determine binding affinity and stoichiometry between IDH3G and its protein partners under controlled conditions.
2. Antibody Development and Validation
The high-purity recombinant IDH3G protein appears well-suited as an antigen for generating specific antibodies against the gamma subunit of mitochondrial isocitrate dehydrogenase. Scientists can use this protein to immunize animals for polyclonal antibody production or as a screening antigen for developing monoclonal antibodies. The purified protein is also likely valuable for validating antibody specificity through Western blot, ELISA, and other immunoassays—helping ensure that developed antibodies specifically recognize IDH3G without cross-reacting to other IDH isoforms.
3. Structural and Biophysical Characterization
This recombinant protein provides material for detailed structural studies of the IDH3G subunit using techniques such as X-ray crystallography, NMR spectroscopy, or cryo-electron microscopy. The purified protein should enable biophysical characterization including thermal stability analysis, circular dichroism spectroscopy to assess secondary structure, and dynamic light scattering to evaluate protein aggregation states. Such studies may provide insights into the structural role of the gamma subunit within the IDH3 complex and how it contributes to overall enzyme architecture.
4. Biochemical Assays for Complex Assembly
The N-terminal His-SUMO tagged IDH3G can be immobilized on affinity matrices to study how the IDH3 complex assembles sequentially by adding purified IDH3A and IDH3B subunits. This approach allows researchers to investigate the order of subunit association and identify critical residues required for complex formation. Scientists might also study how post-translational modifications affect subunit interactions. The tagged protein makes it easier to purify assembled complexes for further biochemical analysis and characterization of the reconstituted enzyme activity.
