BC2LA from Burkholderia cenocepacia linked to Biotin

BC2LA from Burkholderia cenocepacia linked to Biotin

BC2L-A from Burkholderia cenocepacia linked to Biotin is a biotinylated bacterial lectin used for studying mannose-specific carbohydrate interactions, bacterial adhesion, and biofilm formation. This conjugate enables precise detection and quantification of BC2L-A activity in experimental systems utilizing biotin-streptavidin-based assays.

Key Characteristics of BC2L-A:

• Function:
  BC2L-A is a mannose-specific lectin that plays a significant role in bacterial adhesion and biofilm stabilization. It binds to mannose-containing glycans on host tissues and bacterial surfaces, contributing to the structural integrity of biofilms and facilitating colonization during infections such as those in cystic fibrosis patients.
• Structural Features:
  • Composed of a LecB-like domain that associates as a dimer.
  • Displays high specificity for mannose and oligomannose-type N-glycans, critical for its role in bacterial aggregation and biofilm formation.
  • Calcium-dependent binding enhances its structural stability and ligand affinity.
• Biotin Conjugation:
  Biotin labeling enhances the versatility of BC2L-A in research applications by enabling:
  • Streptavidin-based detection: Facilitates ELISA assays, Western blotting, and pull-down experiments to study BC2L-A interactions with mannose-containing substrates.
  • Quantitative binding studies: Allows measurement of mannose-binding kinetics using biotin-streptavidin systems.
  • Cell imaging: Enables visualization of BC2L-A interactions with host glycans through fluorescence microscopy when paired with streptavidin-fluorophore conjugates.

Applications in Research:

1. Biofilm Formation Studies:
   BC2L-A stabilizes biofilms by cross-linking bacterial extracellular polysaccharides (EPS). Biotinylated BC2L-A aids in visualizing biofilm matrix organization and studying inhibitors targeting biofilm formation.
2. Host-Pathogen Interaction Analysis:
   Tracks BC2L-A-mediated adhesion to mannose-containing glycoproteins on epithelial cells, elucidating mechanisms of bacterial colonization.
3. Drug Discovery:
   Screens inhibitors targeting BC2L-A’s mannose-binding sites to disrupt bacterial adhesion or biofilm formation processes. Biotinylated BC2L-A is particularly useful for high-throughput assays assessing anti-biofilm agents.
4. Quorum Sensing Studies:
   Investigates the role of BC2L-A in quorum sensing-regulated processes such as biofilm development and virulence factor expression.

Production and Validation:

• Recombinantly expressed in E. coli and purified via affinity chromatography.
• Biotin labeling validated through binding assays with mannose-containing ligands and functional studies confirming its role in biofilm stabilization and adhesion.

This tool is essential for advancing research into Burkholderia cenocepacia infections, particularly in understanding its virulence mechanisms and developing therapeutic strategies targeting lectin-mediated processes.

Citations:

1. https://pubmed.ncbi.nlm.nih.gov/21909279/
2. https://pmc.ncbi.nlm.nih.gov/articles/PMC3164656/
3. https://en.wikipedia.org/wiki/index.html?curid=8235178
4. https://journals.plos.org/plosone/article?id=10.1371%2Fjournal.pone.0067629