Binding of cholera toxin B subunit to intestinal epithelial cells

We have prepared 125I-labeled cholera toxin B subunit (125I-labeled CT-B, a specific activity of 98Ci/mmol) and found that it binds to rat IEC-6 and human Caco-2 intestinal epithelial cells with high affinity (Kd 3.6 and 3.7nM, respectively). The binding of labeled protein was completely inhibited by unlabeled thymosin-α1 (TM-α1), interferon-α2 (IFN-α2), and the synthetic peptide LKEKK that corresponds to residues 16–20 in TM-α1 and 131–135 in IFN-α2, but was not inhibited by the synthetic peptide KKEKL with inverted amino acid sequence (Ki>10μM). Thus, TM-α1, IFN-α2, and the peptide: LKEKK bind with high affinity and specificity to the cholera toxin receptor on IEC-6 and Caco-2 cells. It was found that CT-B and the peptide: LKEKK at concentrations of 10–1000nM increased in a dose-dependent manner the nitric oxide production and the soluble guanylate cyclase activity in IEC-6 and Caco-2 cells. [Display omitted] •TM-α1, INF-α2, and peptide: LKEKK bind to cholera toxin receptor on intestinal cells.•Residues 16–20 in TM-α1 and 131–135 in IFN-α2 are involved in binding to the receptor.•Binding to the receptor leads to an increase in activity of soluble guanylate cyclase.