Recombinant Arabidopsis thaliana Flavonol 3-O-glucosyltransferase UGT71C1 is expressed in E. coli and includes the full-length protein (1-481 amino acids) with an N-terminal 10xHis-tag for ease of purification. The product demonstrates a purity level greater than 85% as verified by SDS-PAGE. This preparation is intended for research use only, ensuring reliable results for scientific investigations.
Flavonol 3-O-glucosyltransferase UGT71C1 appears to be an enzyme involved in the biosynthesis of flavonoids, a group of plant secondary metabolites. These compounds play essential roles in plant development and defense. UGT71C1 carries out the glucosylation process, which likely influences the solubility and stability of flavonoids. This makes it a valuable tool for researchers studying plant metabolic pathways and their regulation.
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. In Vitro Enzyme Activity Characterization
This recombinant UGT71C1 protein can be used to establish and optimize enzyme activity assays for flavonol 3-O-glucosyltransferase function. Researchers may test various flavonol substrates and UDP-glucose cofactors to determine kinetic parameters such as Km and Vmax values. The N-terminal His-tag simplifies protein purification and quantification for standardized activity measurements. Such experiments could help establish baseline enzymatic properties and substrate specificity profiles for this plant glycosyltransferase.
2. Substrate Specificity and Inhibitor Screening Studies
The purified recombinant protein enables systematic screening of potential substrates beyond natural flavonols to map the enzyme's substrate tolerance and specificity. Researchers can test synthetic flavonoid derivatives, alternative sugar donors, and potential competitive inhibitors in controlled in vitro assays. The high purity level (>85%) suggests reliable results without interference from contaminating enzymes. These studies would provide insights into the structural requirements for substrate recognition and catalytic efficiency.
3. Protein-Protein Interaction Studies
The His-tagged UGT71C1 can serve as bait protein in pull-down assays to identify potential protein partners involved in flavonoid biosynthesis pathways. The tag allows immobilization on nickel-affinity matrices for capturing interacting proteins from Arabidopsis protein extracts. This approach may reveal regulatory proteins, metabolic channeling partners, or other enzymes that form complexes with UGT71C1 in plant cells. The full-length protein (1-481aa) preserves native protein domains that might be important for physiological interactions.
4. Antibody Development and Validation
This recombinant protein serves as an ideal antigen for generating specific antibodies against UGT71C1 for research applications. The purified protein can be used to immunize animals for polyclonal antibody production or as a screening antigen for monoclonal antibody development. Subsequently, the same protein preparation can validate antibody specificity through Western blotting, ELISA, and other immunoassays. The His-tag also allows capture ELISA formats for antibody characterization and quantification studies.
5. Structural and Biophysical Analysis
The recombinant UGT71C1 protein provides material for biophysical characterization studies including circular dichroism spectroscopy, dynamic light scattering, and thermal stability analysis. Researchers can investigate protein folding, oligomerization states, and conformational changes upon substrate or cofactor binding. The high expression level achievable in E. coli and good purity make it suitable for techniques requiring substantial protein quantities. Studies like these could complement enzymatic analyses by providing insights into structure-function relationships.
