Recombinant Mouse Enoyl-CoA hydratase, mitochondrial (Echs1) is produced using a baculovirus expression system and comes with an N-terminal 10xHis tag that makes purification and detection more straightforward. The protein covers the complete mature sequence from amino acids 28 to 290. Purity exceeds 85% based on SDS-PAGE analysis, which should provide dependable results for research work.
Enoyl-CoA hydratase, mitochondrial, appears to play a vital role in the beta-oxidation pathway during fatty acid metabolism. The enzyme catalyzes the hydration of enoyl-CoA to 3-hydroxyacyl-CoA—a step that seems essential for converting stored fats into energy the body can actually use. Understanding this enzyme may be key to grasping how metabolic processes and energy balance work, which is likely why it draws so much attention 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.
Mouse Echs1 is a mitochondrial enoyl-CoA hydratase that requires precise folding, proper cofactor binding, and correct oligomerization for its functional activity in fatty acid β-oxidation. The baculovirus-insect cell expression system provides a eukaryotic environment that supports proper protein folding, post-translational modifications, and complex assembly, significantly increasing the probability of correct folding compared to bacterial systems. However, as a mitochondrial matrix protein, Echs1 may require mitochondrial-specific chaperones or processing for optimal activity. The N-terminal 10xHis tag is relatively small but may potentially interfere with the protein's mitochondrial targeting sequence or functional domains. While the baculovirus system offers favorable folding conditions, experimental validation remains essential to confirm functional hydratase activity.
1. Biochemical Characterization and Enzyme Kinetics Studies
This application's reliability depends on proper folding and functional validation. Enoyl-CoA hydratase activity requires precise tertiary structure and oligomerization. If correctly folded and active (verified), the protein is highly suitable for determining kinetic parameters and substrate specificity. If misfolded/inactive (unverified), enzymatic assays will yield negative or misleading results.
2. Antibody Development and Validation
Antibody development primarily relies on antigenic sequence recognition. If correctly folded (verified), the protein excels for generating conformation-sensitive antibodies. If misfolded/unverified, it remains highly suitable for producing antibodies against linear epitopes.
3. Protein-Protein Interaction Studies
This application requires proper folding validation. Mitochondrial fatty acid oxidation complex formation requires precise tertiary and quaternary structure. If correctly folded (verified), the protein is suitable for identifying physiological interaction partners. If misfolded/unverified, there is a high risk of non-specific binding.
4. Comparative Species Analysis and Evolutionary Studies
Meaningful comparative studies require native protein conformation and functional activity. If correctly folded and active (verified), the protein enables valid functional comparisons across species. If misfolded/inactive (unverified), comparative analyses would yield misleading evolutionary insights.
Final Recommendation & Action Plan
The baculovirus expression system provides favorable eukaryotic folding conditions for this mitochondrial enzyme, but experimental validation of structural integrity and enzymatic activity is essential before reliable use in functional studies. Begin with biochemical characterization to assess folding quality through size-exclusion chromatography (oligomerization state), circular dichroism spectroscopy, and validate hydratase activity using enoyl-CoA substrates. Once correct folding and functional activity are verified, proceed cautiously with Applications 1, 3, and 4 for kinetic studies, interaction mapping, and comparative analyses. Application 2 (antibody development) can proceed immediately regardless of folding status. If misfolding or lack of enzymatic activity is detected, limit applications to linear epitope antibody production and basic biophysical characterization, avoiding all functional studies. For reliable Echs1 research, always include appropriate activity controls using validated hydratase assays and consider the protein's mitochondrial context in experimental design.
