BC2LC-Nt from Burkholderia cenocepacia linked to Fluorescein

BC2LC-Nt from Burkholderia cenocepacia linked to Fluorescein

BC2LC-Nt from Burkholderia cenocepacia linked to Fluorescein is a fluorescently labeled lectin domain used to study fucose-mediated host-pathogen interactions and proinflammatory signaling in chronic infections like cystic fibrosis. Derived from the N-terminal domain of the BC2L-C superlectin, this TNF-α-like lectin binds fucosylated glycans and triggers immune responses independently of carbohydrate recognition.

Key Characteristics of BC2LC-Nt:

• Function:
  • Fucose-specific binding: Targets α1-2/3/4-linked fucose in human histo-blood group antigens (e.g., Lewis^x, H-type 1) to mediate bacterial adhesion to epithelial cells.
  • Proinflammatory activity: Stimulates IL-8 secretion in airway cells via a carbohydrate-independent mechanism, amplifying inflammation in B. cenocepacia infections.
  • Structural role: Forms TNF-α-like trimers that contribute to BC2L-C’s hexameric architecture, enabling dual lectin functionality.
• Structural Features:
  • TNF-α-like fold: Unique among bacterial lectins, resembling human TNF-α but retaining fucose-binding specificity.
  • Flexible oligomerization: Integrates into BC2L-C’s hexameric structure, flanking calcium-dependent C-terminal domains.
  • Molecular weight: ~15–18 kDa (monomer), forming stable trimers post-Fluorescein labeling.
• Fluorescein Conjugation:
  Labeling with fluorescein isothiocyanate (FITC) enables:
  • Visualization of fucose-binding sites in host tissues via fluorescence microscopy.
  • Quantification of lectin-glycan interactions using fluorescence polarization or flow cytometry.
  • High-throughput screening of fucose analogs to block bacterial adhesion or inflammation.

Applications in Research:

1. Host-Pathogen Adhesion Studies:
   Tracks BC2LC-Nt binding to fucosylated epitopes on airway epithelia, elucidating mechanisms of cenocepacia colonization.
2. Inflammatory Response Analysis:
   Investigates IL-8 induction in cystic fibrosis models, linking lectin activity to neutrophil recruitment and tissue damage.
3. Anti-Virulence Drug Development:
   Screens inhibitors targeting BC2LC-Nt’s fucose-binding site or TNF-α-like signaling to reduce bacterial persistence.

Production and Validation:

• Recombinantly expressed in E. coli with affinity tags for purification.
• Fluorescein labeling validated via:
  • Binding assays with fucosylated glycans (e.g., Lewis^x).
  • Cell-based assays confirming IL-8 induction in airway epithelial cultures.

This tool bridges structural biology and immunology, offering insights into B. cenocepacia’s dual strategies of glycan recognition and immune evasion.

Meta Description:
“Explore BC2LC-Nt from Burkholderia cenocepacia, a fluorescently labeled TNF-α-like lectin for studying fucose-mediated adhesion and proinflammatory signaling. Ideal for cystic fibrosis research and anti-virulence drug discovery.”

Citations:

1. https://en.wikipedia.org/wiki/index.html?curid=8235178
2. https://pmc.ncbi.nlm.nih.gov/articles/PMC3164656/
3. https://pubmed.ncbi.nlm.nih.gov/21909279/