Interaction of Cholera Toxin B Subunit with Rat Intestinal Epithelial Cells
- Authors: Navolotskaya E.V.1, Sadovnikov V.B.1, Zinchenko D.V.1, Vladimirov V.I.1,2, Zolotarev Y.A.3
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Affiliations:
- Branch of Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry
- Pushchino State Natural Science Institute
- Institute of Molecular Genetics
- Issue: Vol 44, No 4 (2018)
- Pages: 403-407
- Section: Article
- URL: https://journals.rcsi.science/1068-1620/article/view/228964
- DOI: https://doi.org/10.1134/S1068162018030123
- ID: 228964
Cite item
Abstract
We prepared 125I-labeled cholera toxin B subunit (125I-labeled CT-B, specific activity 98 Ci/mmol) and found that its binding to rat IEC-6 intestinal epithelial cells was high-affinity (Kd 1.9 nM). The binding of labeled protein was completely inhibited by unlabeled thymosin-α1 (TM-α1), interferon-α2 (IFN-α2), and synthetic peptide LKEKK, which corresponds to residues 16–20 in TM-α1 and 131–135 in IFN-α2 (Ki 1.2, 0.9, and 1.6 nM, respectively), but was not inhibited by synthetic peptide KKEKL with inverted amino acid sequence (Ki > 10 μM). Thus, TM-α1, IFN-α2, and the LKEKK peptide bind with high affinity and specificity to CT-B receptor on rIEC-6 cells. It was found that CT-B and the LKEKK peptide at concentrations of 10–1000 nM increased nitric oxide production and soluble guanylate cyclase activity in the cells in a dose-dependent manner.
Keywords
About the authors
E. V. Navolotskaya
Branch of Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry
Author for correspondence.
Email: navolotskaya@bibch.ru
Russian Federation, Pushchino, Moscow oblast, 142290
V. B. Sadovnikov
Branch of Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry
Email: navolotskaya@bibch.ru
Russian Federation, Pushchino, Moscow oblast, 142290
D. V. Zinchenko
Branch of Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry
Email: navolotskaya@bibch.ru
Russian Federation, Pushchino, Moscow oblast, 142290
V. I. Vladimirov
Branch of Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry; Pushchino State Natural Science Institute
Email: navolotskaya@bibch.ru
Russian Federation, Pushchino, Moscow oblast, 142290; Pushchino, Moscow oblast, 142290
Y. A. Zolotarev
Institute of Molecular Genetics
Email: navolotskaya@bibch.ru
Russian Federation, Moscow, 123182