Recombinant Ambrosia artemisiifolia Pectate lyase 1 is produced in E. coli and contains the full-length mature protein spanning amino acids 26 to 398. An N-terminal 6xHis-tag is attached to the protein, which helps with purification and detection. SDS-PAGE analysis confirms the product achieves greater than 90% purity, making it suitable for research work.
Pectate lyase 1 appears to play an important role in breaking down pectin, a major building block of plant cell walls. The enzyme works by cutting α-1,4-glycosidic bonds in pectate - a process that seems central to how plant tissues break apart and how disease develops. Scientists may find this protein particularly useful when studying plant-pathogen interactions, agricultural biotechnology, and how enzymes function.
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. Enzyme Kinetics and Biochemical Characterization Studies
Scientists can use this recombinant pectate lyase 1 from Ambrosia artemisiifolia to study how plant pectate lyases work through laboratory substrate breakdown tests. Researchers might determine the best pH, temperature, and salt conditions for enzyme activity using pectin or pectate as starting materials. The high purity (>90%) and N-terminal His-tag make purification and measurement more straightforward for accurate kinetic studies. This work could help reveal how ragweed pectate lyases compare to those found in other plants.
2. Protein-Protein Interaction Studies
The N-terminal 6xHis-tag allows for pull-down experiments to find potential binding partners or helper molecules that work with this pectate lyase. Scientists can attach the recombinant protein to nickel-coated beads and mix it with plant cell extracts or protein collections to grab onto interacting molecules. Mass spectrometry analysis of these captured complexes might uncover new regulatory proteins or enzyme modifiers. This strategy could shed light on the molecular mechanisms that control how pectate lyase modifies plant cell walls.
3. Antibody Development and Immunological Studies
This purified recombinant protein works well as an antigen for creating specific antibodies against Ambrosia artemisiifolia pectate lyase 1 in laboratory animals. The high purity and known protein sequence should give consistent results when producing polyclonal or monoclonal antibodies. Scientists can test these antibodies using ELISA, Western blot, or immunofluorescence methods with the His-tagged recombinant protein serving as a positive control. Such research-grade antibodies would make it possible to detect and locate native pectate lyase in ragweed plant tissues.
4. Structural Biology and Protein Folding Analysis
Researchers can study this recombinant protein's structure using techniques like circular dichroism spectroscopy, dynamic light scattering, and analytical ultracentrifugation to examine protein folding, stability, and whether multiple protein units stick together. X-ray crystallography or NMR studies could reveal the three-dimensional structure of this ragweed pectate lyase. The His-tag helps purify the protein to the high concentrations that structural biology methods typically require. These investigations may provide insights into the molecular shape and folding characteristics unique to Ambrosia artemisiifolia pectate lyases.
