LecB (PA-IIL) from Pseudomonas aeruginosa linked to Biotin

LecB (PA-IIL) from Pseudomonas aeruginosa linked to Biotin

LecB (PA-IIL) from Pseudomonas aeruginosa linked to Biotin is a biotinylated bacterial lectin used for studying fucose-specific carbohydrate interactions, biofilm formation, and host-pathogen adhesion mechanisms. This conjugate enables precise detection and quantification of LecB activity in experimental systems utilizing biotin-streptavidin-based assays.

Key Characteristics of LecB (PA-IIL):

• Function:
  LecB is a fucose-binding lectin that plays a critical role in biofilm formation, pilus biogenesis, and bacterial adhesion to host cells. It binds with high affinity to fucosylated glycans on epithelial surfaces and extracellular matrices, contributing to P. aeruginosa pathogenicity in infections such as cystic fibrosis and chronic lung conditions.
• Structural Features:
  • Tetrameric protein stabilized by calcium ions, with four carbohydrate-binding sites.
  • High binding affinity for L-fucose (KD=1.5×106 M−1K_D = 1.5 \times 10^6 \, \text{M}^{-1}KD=1.5×106M−1) and lower affinity for D-mannose (KD=3.1×102 M−1K_D = 3.1 \times 10^2 \, \text{M}^{-1}KD=3.1×102M−1).
  • Interacts with outer membrane porin OprF, anchoring LecB to the bacterial surface.
• Biotin Conjugation:
  Biotin labeling enhances the versatility of LecB in research applications by enabling:
  • Streptavidin-based detection: Facilitates ELISA assays, Western blotting, and pull-down experiments.
  • Quantitative binding studies: Allows measurement of fucose-binding kinetics using biotin-streptavidin systems.
  • Cell imaging: Enables visualization of LecB interactions with host glycans through fluorescence microscopy when paired with streptavidin-fluorophore conjugates.

Applications in Research:

1. Biofilm Formation Studies:
   LecB stabilizes biofilms by cross-linking bacterial lipopolysaccharides (LPS) and exopolysaccharides (EPS). Biotinylated LecB aids in visualizing biofilm matrix organization and studying inhibitors targeting biofilm formation.
2. Host-Pathogen Interaction Analysis:
   Tracks LecB-mediated adhesion to fucosylated glycoproteins on epithelial cells, elucidating mechanisms of bacterial colonization and immune evasion.
3. Drug Discovery:
   Screens inhibitors targeting LecB’s fucose-binding sites to disrupt biofilm formation or adhesion processes. Biotinylated LecB is particularly useful for high-throughput assays assessing anti-biofilm agents.
4. Immune Modulation Studies:
   Investigates LecB’s role in suppressing immune responses by inhibiting transendothelial migration of immune cells during infection.

Production and Validation:

• Recombinantly expressed in E. coli and purified via affinity chromatography.
• Biotin labeling validated through binding assays with fucosylated 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:

1. https://journals.asm.org/doi/10.1128/iai.00551-06
2. https://pmc.ncbi.nlm.nih.gov/articles/PMC1698087/
3. https://pmc.ncbi.nlm.nih.gov/articles/PMC6873108/
4. https://journals.plos.org/plosone/article?id=10.1371%2Fjournal.pone.0046857
5. https://pmc.ncbi.nlm.nih.gov/articles/PMC10074054/
6. https://pmc.ncbi.nlm.nih.gov/articles/PMC2681743/
7. https://pubmed.ncbi.nlm.nih.gov/10497867/