LecA (PA-IL) from Pseudomonas aeruginosa linked to Biotin

LecA (PA-IL) from Pseudomonas aeruginosa linked to Biotin LecA (PA-IL) from Pseudomonas aeruginosa linked to Biotin is a biotinylated bacterial lectin used for studying galactose-specific carbohydrate interactions, bacterial adhesion, and host-pathogen interactions. This conjugate enables precise detection and quantification of LecA activity in experimental systems utilizing biotin-streptavidin-based assays. Key Characteristics of LecA (PA-IL): Function : LecA is a tetrameric, galactose-binding lectin that functions as an adhesin and cytotoxin for respiratory epithelial cells. It plays a crucial role in P. aeruginosa pathogenicity by facilitating bacterial adhesion to host cells and contributing to biofilm formation. LecA also interacts with the ABO(H) and P blood group glycosphingolipid antigens, which may enhance tissue infectivity. Structural Features : Consists of four 12.75 kDa subunits, forming a stable tetrameric structure. Exhibits specificity for α-galactose residues, which are critical for its role in bacterial adhesion and biofilm stabilization. The gene encoding LecA, lecA , is regulated by quorum sensing systems involving LasR and RhlR transcriptional regulators. Biotin Conjugation : Biotin labeling enhances the versatility of LecA in research applications by enabling: Streptavidin-based detection : Facilitates ELISA assays, Western blotting, and pull-down experiments to study LecA interactions with galactose-containing substrates. Quantitative binding studies : Allows measurement of galactose-binding kinetics using biotin-streptavidin systems. Cell imaging : Enables visualization of LecA interactions with host glycans through fluorescence microscopy when paired with streptavidin-fluorophore conjugates. Applications in Research: Host-Pathogen Interaction Analysis : Tracks LecA-mediated adhesion to galactose-containing glycoproteins on epithelial cells, elucidating mechanisms of bacterial colonization and immune evasion. Biofilm Formation Studies : LecA stabilizes biofilms by cross-linking bacterial lipopolysaccharides (LPS) and exopolysaccharides (EPS). Biotinylated LecA aids in visualizing biofilm matrix organization and studying inhibitors targeting biofilm formation. Drug Discovery : Screens inhibitors targeting LecA’s galactose-binding sites to disrupt bacterial adhesion or biofilm formation processes. Biotinylated LecA is particularly useful for high-throughput assays assessing anti-biofilm agents. Production and Validation: Recombinantly expressed in E. coli and purified via affinity chromatography. Biotin labeling validated through binding assays with galactose-containing ligands and functional studies confirming its role in biofilm stabilization and adhesion. This tool is essential for advancing research into Pseudomonas aeruginosa infections, particularly in understanding its virulence mechanisms and developing therapeutic strategies targeting lectin-mediated processes. Citations: https://pmc.ncbi.nlm.nih.gov/articles/PMC94786/ https://pubs.acs.org/doi/10.1021/acs.biomac.1c00222 https://www.elicityl-oligotech.com/biotin/577-leca-pa-il-from-pseudomonas-aeruginosa-linked-to-biotin---bacterial-protein-produced-in-e--coli.html https://pubmed.ncbi.nlm.nih.gov/27209461/ https://pubs.acs.org/doi/10.1021/ac4040554