Recombinant Arabidopsis thaliana Alcohol dehydrogenase class-3 (ADH2) is produced in E. coli and includes the complete mature protein sequence from amino acids 2-379. The protein carries an N-terminal 6xHis-tag, which makes purification straightforward. SDS-PAGE analysis shows the product achieves over 90% purity—a level that appears suitable for research applications requiring high-quality protein preparations.
Alcohol dehydrogenase class-3 (ADH2) from Arabidopsis thaliana catalyzes the conversion of alcohols to aldehydes, a reaction that's important in cellular metabolism. This enzyme likely participates in detoxification processes and helps plants adapt metabolically to stress conditions. For researchers working on plant biology, ADH2 may provide useful insights into enzymatic activity and how regulatory mechanisms function in plants.
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. Biochemical Characterization of Plant Alcohol Dehydrogenase Activity
Researchers can use this recombinant ADH2 protein to set up in vitro enzyme assays and characterize the biochemical properties of Arabidopsis alcohol dehydrogenase class-3. These studies would determine which substrates the enzyme prefers and establish kinetic parameters like Km and Vmax using different alcohol substrates along with NAD+/NADH cofactors. The purified protein allows for systematic testing of pH and temperature conditions to better understand how the enzyme works catalytically. The N-terminal 6xHis tag makes purification easier and allows the protein to be immobilized for repeated experiments.
2. Comparative Enzyme Evolution Studies
ADH2 could serve as a reference point for comparative studies looking at how alcohol dehydrogenases have evolved across different plant species. Scientists might compare catalytic efficiency, substrate preferences, and structural features between Arabidopsis ADH2 and similar enzymes from other plants. This type of work supports phylogenetic analyses and may help identify which functional domains remain conserved versus those that have diverged within the alcohol dehydrogenase class-3 family. Using the standardized E. coli expression system should provide consistent enzyme preparations for fair comparisons.
3. Antibody Development and Validation
This purified recombinant protein appears well-suited as an antigen for generating antibodies specific to Arabidopsis ADH2. The high purity level (>90%) and full-length mature protein sequence should ensure proper epitope presentation during immunization protocols. Researchers can then validate any generated antibodies using the recombinant protein in Western blots, ELISA assays, and immunoprecipitation experiments. The N-terminal His-tag makes it easy to capture and present the antigen in different immunoassay formats.
4. Protein-Protein Interaction Studies
The His-tagged recombinant ADH2 might be useful in pull-down assays to identify potential protein partners from Arabidopsis protein extracts. Scientists could immobilize the protein on nickel-affinity resins to capture any interacting proteins, then identify these by mass spectrometry. This approach may help reveal the cellular networks and regulatory mechanisms that involve alcohol dehydrogenase in plant metabolism. The recombinant protein also works as a positive control in yeast two-hybrid screens targeting ADH2 interactions.
5. Structural and Biophysical Analysis
This recombinant protein provides the material needed for detailed structural studies. Techniques like X-ray crystallography, NMR spectroscopy, and cryo-electron microscopy could reveal the three-dimensional structure of plant alcohol dehydrogenase class-3. The purified protein enables biophysical characterization through dynamic light scattering, circular dichroism spectroscopy, and thermal stability analysis. Such studies contribute to understanding how structure relates to function and might guide future protein engineering efforts. The consistent E. coli expression system should ensure reproducible protein preparations for structural work.
