Effects of Claudin-1 on the Action of Clostridium perfringens Enterotoxin in Caco-2 Cells

Iman Mehdizadeh Gohari, Jihong Li, Mauricio Navarro, Francisco Uzal, Bruce McClane

Research output: Contribution to journalArticle

Abstract

Clostridium perfringens enterotoxin (CPE) contributes to diarrhea and an often-lethal enterotoxemia. CPE action starts when it binds to claudin receptors, forming a small complex (90 kDa). Six small complexes then oligomerize to create prepores, followed by insertion of beta-hairpins from CPE to form beta-barrel pores named CH-1 or CH-2. Of the ~27 members of the human claudin protein family, only some bind CPE. However, both receptor claudins and the nonreceptor claudin-1 (CLDN-1) are associated with the small and CH-1/CH-2 CPE complexes. Therefore, this study evaluated whether claudin-1 affects CPE action by generating a CLDN-1 null mutant in Caco-2 cells using CRISPR-Cas9. Compared to wild-type Caco-2 cells, paracellular permeability of the CLDN-1 mutant was significantly enhanced, suggesting that claudin-1 may reduce CPE absorption during enterotoxemia. The CLDN-1 mutant was also markedly more sensitive than wild-type Caco-2 cells to apically-applied CPE. The mechanism behind this increased sensitivity involved higher CPE binding by the CLDN-1 mutant vs. wild-type Caco-2 cells, which led to more CH-1/CH-2 complex formation. However, the CH-1/CH-2 complexes formed by the CLDN-1 mutant were less stable or trypsin resistant than those of wild-type cells. These results indicate that, although a nonreceptor, CLDN-1 positively and negatively influences CPE action.

Original languageEnglish (US)
JournalToxins
Volume11
Issue number10
DOIs
StatePublished - Oct 9 2019

Keywords

  • claudins
  • Clostridium perfringens
  • diarrheal disease
  • enterotoxin
  • occludin
  • tight junctions

ASJC Scopus subject areas

  • Toxicology
  • Health, Toxicology and Mutagenesis

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