Recombinant Ambrosia artemisiifolia Pectate lyase 5 gets expressed in a yeast system and includes a full-length mature protein corresponding to amino acids 26-396. The protein is engineered with an N-terminal 6xHis tag, which makes purification and detection more straightforward. It shows purity greater than 90% as verified by SDS-PAGE, likely ensuring reliable performance for research applications.
Pectate lyase 5 from Ambrosia artemisiifolia is an enzyme involved in the breakdown of pectin, a major component of the plant cell wall. This protein appears to play a crucial role in plant tissue degradation and may be of interest in research focused on plant pathology and allergen studies, given the importance of its source species, short ragweed, in allergenic responses.
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
This recombinant pectate lyase 5 from Ambrosia artemisiifolia can be used to investigate the enzymatic properties and kinetic parameters of plant pectate lyases through in vitro assays. Researchers might determine substrate specificity, optimal pH and temperature conditions, and catalytic efficiency using various pectate substrates. The high purity (>90%) and N-terminal His-tag make purification and handling easier for detailed biochemical analysis. Such studies would likely contribute to understanding the role of pectate lyases in plant cell wall modification and ragweed biology.
2. Comparative Enzyme Evolution and Structure-Function Analysis
The protein could serve as a valuable tool for comparative studies examining pectate lyase evolution across different plant species, particularly within the Asteraceae family. Researchers might compare its enzymatic properties, substrate preferences, and structural features with pectate lyases from other plants to understand evolutionary adaptations. The recombinant protein allows controlled comparative experiments that would be difficult to perform with native plant extracts. This application may support broader research into plant cell wall enzyme diversity and functional specialization.
3. His-Tag Affinity Purification and Protein Interaction Studies
The N-terminal 6xHis-tag allows for straightforward purification using nickel affinity chromatography and makes pull-down experiments possible to identify potential protein interaction partners. Researchers can use this tagged protein in binding assays with plant cell wall components or other proteins involved in cell wall metabolism. The tag also allows for easy detection and quantification in various experimental setups, making it suitable for studying protein-protein or protein-substrate interactions in controlled in vitro systems.
4. Antibody Development and Immunological Research
This highly purified recombinant protein may serve as an excellent antigen for generating specific antibodies against Ambrosia artemisiifolia pectate lyase 5. The resulting antibodies could be used in various research applications including Western blotting, immunohistochemistry, and ELISA-based detection systems for studying ragweed biology. The recombinant nature ensures consistent antigen quality and availability for immunization protocols, while the high purity appears to minimize cross-reactivity with other proteins during antibody development.
