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Identification of claudin-4 binder that attenuates tight junction barrier function by TR-FRET-based screening assay
Claudins are key functional and structural components of tight junctions (TJs) in epithelial cell sheets. The C-terminal fragment of Clostridium perfringens enterotoxin (C-CPE) binds to claudin-4 and reversibly modulates intestinal TJ seals, thereby enhancing paracellular transport of solutes. Howev...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5674027/ https://www.ncbi.nlm.nih.gov/pubmed/29109448 http://dx.doi.org/10.1038/s41598-017-15108-y |
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author | Watari, Akihiro Kodaka, Miki Matsuhisa, Koji Sakamoto, Yuta Hisaie, Kota Kawashita, Norihito Takagi, Tatsuya Yamagishi, Yoshiaki Suzuki, Hidehiko Tsujino, Hirofumi Yagi, Kiyohito Kondoh, Masuo |
author_facet | Watari, Akihiro Kodaka, Miki Matsuhisa, Koji Sakamoto, Yuta Hisaie, Kota Kawashita, Norihito Takagi, Tatsuya Yamagishi, Yoshiaki Suzuki, Hidehiko Tsujino, Hirofumi Yagi, Kiyohito Kondoh, Masuo |
author_sort | Watari, Akihiro |
collection | PubMed |
description | Claudins are key functional and structural components of tight junctions (TJs) in epithelial cell sheets. The C-terminal fragment of Clostridium perfringens enterotoxin (C-CPE) binds to claudin-4 and reversibly modulates intestinal TJ seals, thereby enhancing paracellular transport of solutes. However, the use of C-CPE as an absorption enhancer is limited by the molecule’s immunogenicity and manufacturing cost. Here, we developed a high-throughput screening system based on the Time-Resolved Fluorescence Resonance Energy Transfer (TR-FRET) method to identify claudin-4 binders in a library collection of 32,560 compounds. Thiostrepton, identified from the screen, decreased transepithelial electrical resistance and increased flux of 4-kDa fluorescein isothiocyanate–labelled dextran (FD-4) in Caco-2 cell monolayers, a model of intestinal epithelium. Thiostrepton changed the expression, but not the localisation, of TJ components. Treatment of rat jejunum with thiostrepton increased the absorption of FD-4 without tissue toxicity, indicating that thiostrepton is a novel claudin-4 binder that enhances intestinal permeability. The screening system may therefore be a useful tool for identifying claudin-4 binders to enhance drug absorption in mucosa. |
format | Online Article Text |
id | pubmed-5674027 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-56740272017-11-15 Identification of claudin-4 binder that attenuates tight junction barrier function by TR-FRET-based screening assay Watari, Akihiro Kodaka, Miki Matsuhisa, Koji Sakamoto, Yuta Hisaie, Kota Kawashita, Norihito Takagi, Tatsuya Yamagishi, Yoshiaki Suzuki, Hidehiko Tsujino, Hirofumi Yagi, Kiyohito Kondoh, Masuo Sci Rep Article Claudins are key functional and structural components of tight junctions (TJs) in epithelial cell sheets. The C-terminal fragment of Clostridium perfringens enterotoxin (C-CPE) binds to claudin-4 and reversibly modulates intestinal TJ seals, thereby enhancing paracellular transport of solutes. However, the use of C-CPE as an absorption enhancer is limited by the molecule’s immunogenicity and manufacturing cost. Here, we developed a high-throughput screening system based on the Time-Resolved Fluorescence Resonance Energy Transfer (TR-FRET) method to identify claudin-4 binders in a library collection of 32,560 compounds. Thiostrepton, identified from the screen, decreased transepithelial electrical resistance and increased flux of 4-kDa fluorescein isothiocyanate–labelled dextran (FD-4) in Caco-2 cell monolayers, a model of intestinal epithelium. Thiostrepton changed the expression, but not the localisation, of TJ components. Treatment of rat jejunum with thiostrepton increased the absorption of FD-4 without tissue toxicity, indicating that thiostrepton is a novel claudin-4 binder that enhances intestinal permeability. The screening system may therefore be a useful tool for identifying claudin-4 binders to enhance drug absorption in mucosa. Nature Publishing Group UK 2017-11-06 /pmc/articles/PMC5674027/ /pubmed/29109448 http://dx.doi.org/10.1038/s41598-017-15108-y Text en © The Author(s) 2017 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Watari, Akihiro Kodaka, Miki Matsuhisa, Koji Sakamoto, Yuta Hisaie, Kota Kawashita, Norihito Takagi, Tatsuya Yamagishi, Yoshiaki Suzuki, Hidehiko Tsujino, Hirofumi Yagi, Kiyohito Kondoh, Masuo Identification of claudin-4 binder that attenuates tight junction barrier function by TR-FRET-based screening assay |
title | Identification of claudin-4 binder that attenuates tight junction barrier function by TR-FRET-based screening assay |
title_full | Identification of claudin-4 binder that attenuates tight junction barrier function by TR-FRET-based screening assay |
title_fullStr | Identification of claudin-4 binder that attenuates tight junction barrier function by TR-FRET-based screening assay |
title_full_unstemmed | Identification of claudin-4 binder that attenuates tight junction barrier function by TR-FRET-based screening assay |
title_short | Identification of claudin-4 binder that attenuates tight junction barrier function by TR-FRET-based screening assay |
title_sort | identification of claudin-4 binder that attenuates tight junction barrier function by tr-fret-based screening assay |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5674027/ https://www.ncbi.nlm.nih.gov/pubmed/29109448 http://dx.doi.org/10.1038/s41598-017-15108-y |
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