Mechanism of Clostridium perfringens Enterotoxin Interaction with Claudin-3/-4 Protein Suggests Structural Modifications of the Toxin to Target Specific Claudins

Veshnyakova, Anna; Piontek, Jörg; Protze, Jonas; Waziri, Negar; Heise, Ivonne; Krause, Gerd

doi:10.1074/jbc.m111.312165

Cited by 79 publications

(104 citation statements)

References 37 publications

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“…Luminal proteases such as cap1 may contribute to such dynamic behavior of TJ or rely upon the opening event to gain access to its claudin targets. The trans claudin interaction depends upon its ECL2 domain, which consists of ∼25 amino acids with a helixturn-helix motif (35) and also mediates claudin interactions with extracellular pathogens such as the Clostridium perfringens enterotoxin (51). Mutagenic screening in claudin-5 has identified three loci of amino acids in ECL2 (F147, Y148, and Y158) critical for its trans interaction (35).…”

Section: Discussionmentioning

confidence: 99%

The Cap1–claudin-4 regulatory pathway is important for renal chloride reabsorption and blood pressure regulation

Gong

Yang³

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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The paracellular pathway through the tight junction provides an important route for transepithelial chloride reabsorption in the kidney, which regulates extracellular salt content and blood pressure. Defects in paracellular chloride reabsorption may in theory cause deregulation of blood pressure. However, there is no evidence to prove this theory or to demonstrate the in vivo role of the paracellular pathway in renal chloride handling. Here, using a tissue-specific KO approach, we have revealed a chloride transport pathway in the kidney that requires the tight junction molecule claudin-4. The collecting duct-specific claudin-4 KO animals developed hypotension, hypochloremia, and metabolic alkalosis due to profound renal wasting of chloride. The claudin-4-mediated chloride conductance can be regulated endogenously by a proteasechannel-activating protease 1 (cap1). Mechanistically, cap1 regulates claudin-4 intercellular interaction and membrane stability. A putative cap1 cleavage site has been identified in the second extracellular loop of claudin-4, mutation of which abolished its regulation by cap1. The cap1 effects on paracellular chloride permeation can be extended to other proteases such as trypsin, suggesting a general mechanism may also exist for proteases to regulate the tight junction permeabilities. Together, we have discovered a theory that paracellular chloride permeability is physiologically regulated and essential to renal salt homeostasis and blood pressure control.chloride channel | epithelium | hypertension C hloride is the most abundant extracellular anion and thereby determines extracellular fluid volume (ECFV) and blood pressure (BP) (1, 2). The kidney plays a vital role in ECFV and BP control through complex regulatory mechanisms acting upon ion channels and transporters located in the aldosterone-sensitive distal nephron (ASDN) (3, 4). The ASDN comprises the distal convoluted tubule (DCT), the connecting tubule (CNT), and the collecting duct (CD). The primary chloride transport mechanism in the DCT is through the thiazide-sensitive Na + /Cl − cotransporter (NCC) to reabsorb Cl − coupled with equal moles of Na + (5). The chloride transport mechanism in the CNT and CD, on the other hand, has been under debate for many years. Recent advances have identified an electroneutral transport pathway for chloride using the Cl − /bicarbonate exchanger (Slc26a4; pendrin) (6) and the Na + -driven Cl − /bicarbonate exchanger (Slc4a8; NDCBE) (7) in the β-intercalated cells (ICs) of CNT and CD. However, such an electroneutral pathway is not able to couple Cl − reabsorption with epithelial sodium channel (ENaC)-based Na + reabsorption that takes place in the principal cells (PCs) of CNT and CD (8), despite many efforts to connect ENaC and pendrin gene expression through endocrine or paracrine mechanisms (9, 10). We and others have previously demonstrated the presence of a paracellular Cl − pathway or "chloride shunt" in vitro in the CNT and CD cells (11,12). The paracellular Cl − channel is made of a key claud...

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Section: Discussionmentioning

confidence: 99%

The Cap1–claudin-4 regulatory pathway is important for renal chloride reabsorption and blood pressure regulation

Gong

Yang³

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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“…CPE action begins by binding of the toxin to its receptors, which include certain members of the claudin tight junction protein family (62)(63)(64)(65)(66)(67)(68). Claudins are ϳ20-to 25-kDa proteins that consist of four transmembrane domains and two extracellular loops (ECLs) (69,70).…”

Section: Toxins That Can Be Either Chromosomally or Plasmid Encodedmentioning

confidence: 99%

“…CPE binds, via a pocket on its C-terminal domain, to the second ECL of claudin receptors (71). Particularly important for this receptor binding interaction are (i) an Asn residue located near the middle of ECL2 on receptor claudins and (ii) Tyr residues present at amino acids 306, 310, and 312 in the CPE C terminus (1,65,67,72). Pore former a C, chromosomal; P, plasmid (13-16, 18, 23, 25-27).…”

Section: Toxins That Can Be Either Chromosomally or Plasmid Encodedmentioning

confidence: 99%

Toxin Plasmids of Clostridium perfringens

Adams

Bannam

et al. 2013

Microbiol Mol Biol Rev

225

282

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SUMMARY In both humans and animals, Clostridium perfringens is an important cause of histotoxic infections and diseases originating in the intestines, such as enteritis and enterotoxemia. The virulence of this Gram-positive, anaerobic bacterium is heavily dependent upon its prolific toxin-producing ability. Many of the ∼16 toxins produced by C. perfringens are encoded by large plasmids that range in size from ∼45 kb to ∼140 kb. These plasmid-encoded toxins are often closely associated with mobile elements. A C. perfringens strain can carry up to three different toxin plasmids, with a single plasmid carrying up to three distinct toxin genes. Molecular Koch's postulate analyses have established the importance of several plasmid-encoded toxins when C. perfringens disease strains cause enteritis or enterotoxemias. Many toxin plasmids are closely related, suggesting a common evolutionary origin. In particular, most toxin plasmids and some antibiotic resistance plasmids of C. perfringens share an ∼35-kb region containing a Tn 916 -related conjugation locus named tcp (transfer of clostridial plasmids). This tcp locus can mediate highly efficient conjugative transfer of these toxin or resistance plasmids. For example, conjugative transfer of a toxin plasmid from an infecting strain to C. perfringens normal intestinal flora strains may help to amplify and prolong an infection. Therefore, the presence of toxin genes on conjugative plasmids, particularly in association with insertion sequences that may mobilize these toxin genes, likely provides C. perfringens with considerable virulence plasticity and adaptability when it causes diseases originating in the gastrointestinal tract.

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“…Known CPE receptors include certain members of the claudin family of tight junction proteins (7,8,18,25,36). Binding of the toxin to claudins initially results in formation of an SDS-sensitive, small (ϳ90-kDa) complex (38).…”

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confidence: 99%

Cysteine-Scanning Mutagenesis Supports the Importance of Clostridium perfringens Enterotoxin Amino Acids 80 to 106 for Membrane Insertion and Pore Formation

et al. 2012

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Clostridium perfringens enterotoxin (CPE) causes the gastrointestinal symptoms of the second most common bacterial foodborne illness. Previous studies suggested that a region named TM1, which has amphipathic characteristics and spans from amino acids 81 to 106 of the native CPE protein, forms a ␤-hairpin involved in ␤-barrel pore formation. To further explore the potential role of TM1 in pore formation, the single Cys naturally present in CPE at residue 186 was first altered to alanine by mutagenesis; the resultant rCPE variant, named C186A, was shown to retain cytotoxic properties. Cys-scanning mutagenesis was then performed in which individual Cys mutations were introduced into each TM1 residue of the C186A variant. When those Cys variants were characterized, three variants were identified that exhibit reduced cytotoxicity despite possessing binding and oligomerization abilities similar to those of the C186A variant from which they were derived. Pronase challenge experiments suggested that the reduced cytotoxicity of those two Cys variants, i.e., the F91C and F95C variants, which model to the tip of the ␤-hairpin, was attributable to a lessened ability of these variants to insert into membranes after oligomerization. In contrast, another Cys variant, i.e., the G103C variant, with impaired cytotoxicity apparently inserted into membranes after oligomerization but could not form a pore with a fully functional channel. Collectively, these results support the TM1 region forming a ␤-hairpin as an important step in CPE insertion and pore formation. Furthermore, this work identifies the first amino acid residues specifically involved in those two steps in CPE action.

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Mechanism of Clostridium perfringens Enterotoxin Interaction with Claudin-3/-4 Protein Suggests Structural Modifications of the Toxin to Target Specific Claudins

Cited by 79 publications

References 37 publications

The Cap1–claudin-4 regulatory pathway is important for renal chloride reabsorption and blood pressure regulation

The Cap1–claudin-4 regulatory pathway is important for renal chloride reabsorption and blood pressure regulation

Toxin Plasmids of Clostridium perfringens

Cysteine-Scanning Mutagenesis Supports the Importance of Clostridium perfringens Enterotoxin Amino Acids 80 to 106 for Membrane Insertion and Pore Formation

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