Recombinant Human 3-hydroxyacyl-CoA dehydrogenase type-2 (HSD17B10) is expressed in E. coli and contains the complete mature protein sequence covering amino acids 2-261. An N-terminal 6xHis-SUMO tag is attached to help with purification and stability. SDS-PAGE analysis shows the product achieves greater than 90% purity, which appears to make it reliable for research work.
HSD17B10 protein serves an important function in mitochondrial pathways. It's involved in fatty acid oxidation and steroid metabolism processes. The enzyme is essential for breaking down various substrates - this includes branched-chain amino acids and neuroactive steroids. Given its central role in metabolic pathways, the protein has become a key target for biochemical and medical research, particularly for studies examining mitochondrial function and metabolic disorders.
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.
Based on the provided information, the recombinant human HSD17B10 is expressed in E. coli, a prokaryotic system that is generally suitable for producing soluble enzymatic proteins like HSD17B10. As a mitochondrial dehydrogenase, HSD17B10 is a relatively small (261aa) soluble enzyme that does not require complex post-translational modifications for its catalytic activity. The protein is expressed as the full-length mature form (2-261aa) with an N-terminal 6xHis-SUMO tag and >90% purity. The SUMO tag may enhance solubility and proper folding. However, since activity is explicitly unverified and the protein requires precise folding for its dehydrogenase activity and NAD+ cofactor binding, the protein cannot be assumed to be correctly folded or bioactive without experimental validation of its enzymatic activity using appropriate substrates.
1. Protein-Protein Interaction Studies Using His-Tag Affinity Purification
The N-terminal 6xHis-SUMO tag enables technical feasibility for pull-down assays. However, if HSD17B10 is misfolded, it may not interact physiologically with true binding partners. This dehydrogenase forms a homotetramer and requires proper oligomerization for some interactions. This application should only be pursued after confirming proper folding and tetramer formation through biophysical characterization.
2. Antibody Development and Validation
The recombinant HSD17B10 can serve as an effective immunogen for generating antibodies that recognize linear epitopes. The full-length sequence ensures comprehensive epitope coverage. Validation against native HSD17B10 from mammalian mitochondria is recommended to ensure physiological relevance.
3. Biochemical Characterization and Enzyme Kinetics Studies
Basic biochemical characterization is feasible, but enzyme kinetics studies require activity validation first. If HSD17B10 is misfolded, kinetic parameters (Km, Vmax) will be invalid. These studies should only follow confirmation of dehydrogenase activity using known substrates (e.g., 3-hydroxyacyl-CoA derivatives).
4. Structural and Biophysical Analysis
This application is well-suited for initial characterization. Techniques like circular dichroism spectroscopy, analytical ultracentrifugation, and size-exclusion chromatography can assess folding state and oligomeric structure. These studies are valuable even before activity validation.
Final Recommendation & Action Plan
Given that HSD17B10 is a soluble enzyme expressed in a prokaryotic system with a solubility-enhancing SUMO tag, the probability of proper folding is relatively high. However, recommend first performing: 1) Functional validation using dehydrogenase activity assays with 3-hydroxyacyl-CoA substrates and NAD+ cofactor; 2) Biophysical characterization (size-exclusion chromatography with multi-angle light scattering) to confirm proper tetramer formation; 3) If possible, SUMO tag removal and comparison with tag-cleaved protein. Antibody development can proceed immediately. Avoid interaction studies until proper folding and oligomerization are confirmed. For reliable kinetic studies, use activity-verified protein and include appropriate controls.
