Recombinant Human Fructose-bisphosphate aldolase C (ALDOC) is produced through a baculovirus expression system, creating a full-length protein that spans amino acids 1-364. The protein carries a C-terminal 6xHis tag, which streamlines both purification and detection processes. SDS-PAGE analysis indicates a purity level greater than 90%, suggesting this protein may serve well for research applications that demand high-quality reagents.
Fructose-bisphosphate aldolase C (ALDOC) functions as an enzyme in the glycolytic pathway. It catalyzes the reversible conversion of fructose-1,6-bisphosphate to glyceraldehyde 3-phosphate and dihydroxyacetone phosphate. The enzyme appears to play a critical role in energy metabolism and shows predominant expression in brain tissue. Research into ALDOC is likely significant for understanding metabolic processes and may help investigate various neurological conditions.
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.
Human ALDOC is a glycolytic enzyme that requires precise folding, oligomerization, and active site formation for its enzymatic activity. The baculovirus-insect cell expression system provides a eukaryotic environment that supports proper protein folding, post-translational modifications, and tetramer formation, significantly increasing the probability of correct folding. The C-terminal 6xHis tag is relatively small and unlikely to interfere with the protein's functional domains or oligomerization interface. While the baculovirus system offers favorable folding conditions, experimental validation remains essential to confirm structural integrity and enzymatic activity.
1. Protein-Protein Interaction Studies Using His-Tag Pull-Down Assays
This application's reliability depends on proper folding validation. ALDOC interactions with other glycolytic enzymes or signaling proteins require precise tertiary and quaternary structure. If correctly folded (verified), the protein is suitable for identifying physiological interaction partners. If misfolded/unverified, there is high risk of non-specific binding or interaction failure, making results biologically misleading.
2. Antibody Development and Validation
Antibody development relies on antigenic sequence recognition, regardless of folding state. If correctly folded (as verified), the protein excels at generating conformation-sensitive antibodies that recognize native ALDOC epitopes. If misfolded/unverified, it remains highly suitable for producing antibodies against linear epitopes, which are still valuable for detection applications.
3. Biochemical Characterization and Enzyme Kinetics Analysis
These studies are essential for determining folding status and functional competence. Functional characterization requires proper folding, but basic biophysical analysis is valuable regardless. If correctly folded and active (verified), characterization provides reliable data on enzymatic activity, kinetic parameters, and oligomerization state. If misfolded/inactive (unverified), biochemical assays will yield negative results, though basic physical properties can be characterized.
4. His-Tagged Protein Purification Method Development
Tag-based purification is independent of protein folding and function. Purification method development depends on tag functionality. The His-tag allows for optimization of nickel-affinity chromatography protocols under various conditions.
Final Recommendation & Action Plan
The baculovirus expression system provides favorable eukaryotic folding conditions for this metabolic enzyme, but experimental validation of structural integrity and enzymatic activity is essential before reliable use in functional studies. Begin with Application 3 (Biochemical Characterization) to assess folding quality through size-exclusion chromatography (tetramer formation), circular dichroism spectroscopy, and validate enzymatic activity using standard aldolase assays with fructose-1,6-bisphosphate as substrate. Once correct folding and functional activity are verified, proceed cautiously with Application 1 for interaction studies. Applications 2 and 4 (antibody development and purification optimization) can proceed immediately regardless of folding status. If misfolding is detected, limit applications to linear epitope antibody production and purification method development, avoiding all functional interaction and enzymatic studies. For reliable ALDOC research, always include appropriate activity controls and consider the protein's tetrameric nature in experimental design.
