Lambda‐Toxin of <i>Clostridium perfringens</i> Activates the Precursor of Epsilon‐Toxin by Releasing Its N‐ and C‐Terminal Peptides

Minami, Junzaburo; Katayama, Seiichi; Matsushita, Osamu; Matsushita, Chieko; Okabe, Akinobu

doi:10.1111/j.1348-0421.1997.tb01888.x

Cited by 157 publications

(157 citation statements)

References 39 publications

(15 reference statements)

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“…ETX is the third most lethal of all clostridial toxins, ranking behind only the Clostridium botulinum and Clostridium tetani neurotoxins [4]. The fact that the 50% lethal dose (LD 50 ) of ETX in mice is 50 ng·kg −1 [5] underpins the potential to use this toxin as a bioterrorist weapon; therefore, ETX is regarded as a potential bioterrorism agent by the US Government Centers for Disease Control and Prevention [3]. …”

Section: Introductionmentioning

confidence: 99%

“…The relatively inactive secreted prototoxin is converted to the fully active mature toxin by proteases of the host, such as trypsin and chymotrypsin [6], or by the C. perfringens λ-protease [5]. Active ETX binds to the intestinal epithelium to induce epithelial permeability in the absence of overt histologic damage [7] and enters the bloodstream.…”

Section: Introductionmentioning

confidence: 99%

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Hemolysis in human erythrocytes by Clostridium perfringens epsilon toxin requires activation of P2 receptors

et al. 2018

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Epsilon-toxin (ETX) is produced by types B and D strains of Clostridium perfringens, which cause fatal enterotoxaemia in sheep, goats and cattle. Previous studies showed that only a restricted number of cell lines are sensitive to ETX and ETX-induced hemolysis has not previously been reported. In this study, the hemolytic ability of ETX was examined using erythrocytes from 10 species including murine, rabbit, sheep, monkey and human. We found that ETX caused hemolysis in human erythrocytes (HC50 = 0.2 μM) but not erythrocytes from the other test species. Moreover, the mechanism of ETX-induced hemolysis was further explored. Recent studies showed that some bacterial toxins induce hemolysis through purinergic receptor (P2) activation. Hence, the function of purinergic receptors in ETX-induced hemolysis was tested, and we found that the non-selective P2 receptor antagonists PPADS inhibited ETX-induced lysis of human erythrocytes in a concentration-dependent manner, indicating that ETX-induced hemolysis requires activation of purinergic receptors. P2 receptors comprise seven P2X (P2X1–7) and eight P2Y (P2Y1, P2Y2, P2Y4, P2Y6, and P2Y11–P2Y14) receptor subtypes. The pattern of responsiveness to more selective P2-antagonists implies that both P2Y13 and P2X7 receptors are involved in ETX-induced hemolysis in human species. Furthermore, we demonstrated that extracellular ATP is likely not involved in ETX-induced hemolysis and the activation of P2 receptors. These findings clarified the mechanism of ETX-induced hemolysis and provided new insight into the activities and ETX mode of action.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Hemolysis in human erythrocytes by Clostridium perfringens epsilon toxin requires activation of P2 receptors

et al. 2018

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“…Another characteristic of ε-toxin is the activation of the inactive precursor (ε-protoxin) by proteases such as trypsin, chymotrypsin (Hunter et al, 1992), and λ-protease produced by C. perfringens. This activation is accompanied by removal of both N-and C-terminal peptides (Jin, et al 1996;Minami et al, 1997).…”

Section: Epsilon-toxin (ε ε ε ε ε)mentioning

confidence: 99%

Large scale purification of Clostridium perfringens toxins: a review

Cavalcanti¹,

Porto²,

Porto³

et al. 2004

Rev. Bras. Cienc. Farm.

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Clostridium perfringens, a Gram-positive anaerobic bacterium, is widespread in the environment and commonly found in the intestines of animals, including humans. C. perfringens strains are classified into five toxinotypes (A, B, C, D and E) based on the production of four major toxins (α, β, ε, ι). However the toxins (theta, delta, lambda and enterotoxin) are also synthesized by C. perfringens strain. Many attempts to purify the toxins produced by C. perfringens have been proposed. In this review we discuss the purification methods used to isolate toxins from C. perfringens reported in last four decades.

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“…This toxin accumulates mainly in the brain and kidneys when intravenously injected into rats (14), causing injury to neuronal cells (15)(16)(17) and cerebral blood vessels (18). One characteristic feature of the toxin is its extraordinarily high potency; 20 ng of ET kills a mouse within 1 h. 2 We have previously shown that proteolytic activation of ⑀-protoxin (ProET) is due to the removal of a C-terminal peptide (19,20) and that activated ET forms a heptamer in rat synaptosomal membranes (20). Similar ET oligomerization has been reported for Madin-Darby canine kidney (MDCK) cell membranes, where the potassium ion permeability is increased by the toxin (21).…”

mentioning

confidence: 99%

Clostridium perfringens ε-Toxin Forms a Heptameric Pore within the Detergent-insoluble Microdomains of Madin-Darby Canine Kidney Cells and Rat Synaptosomes

Miyata

Minami

Tamai

et al. 2002

Journal of Biological Chemistry

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Clostridium perfringens ⑀-toxin, which is responsible for enterotoxaemia in ungulates, forms a heptamer in rat synaptosomal and Madin-Darby canine kidney (MDCK) cell membranes, leading to membrane permealization. Thus, the toxin may target the detergent-resistant membrane domains (DRMs) of these membranes, in analogy to aerolysin, a heptameric pore-forming toxin that associates with DRMs. To test this idea, we examined the distribution of radiolabeled ⑀-toxin in DRM and detergent-soluble membrane fractions of MDCK cells and rat synaptosomal membranes. When MDCK cells and synaptosomal membranes were incubated with the toxin and then fractionated by cold Triton X-100 extraction and flotation on sucrose gradients, the heptameric toxin was detected almost exclusively in DRMs. The results of a toxin overlay assay revealed that the toxin preferentially bound to and heptamerized in the isolated DRMs. Furthermore, cholesterol depletion by methyl-␤-cyclodextrin abrogated their association and lowered the cytotoxicity of the toxin toward MDCK cells. When ⑀-protoxin, an inactive precursor able to bind to but unable to heptamerize in the membrane, was incubated with MDCK cell membranes, it was detected mainly in their DRMs. These results suggest that the toxin is concentrated and induced to heptamerize on binding to a putative receptor located preferentially in DRMs, with all steps from initial binding through pore formation completed within the same DRMs.

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Lambda‐Toxin of Clostridium perfringens Activates the Precursor of Epsilon‐Toxin by Releasing Its N‐ and C‐Terminal Peptides

Abstract: Abstract:The effect of ƒÉ-toxin, a thermolysin-like metalloprotease of Clostridium perfringens, on the inac-

Cited by 157 publications

References 39 publications

Hemolysis in human erythrocytes by Clostridium perfringens epsilon toxin requires activation of P2 receptors

Hemolysis in human erythrocytes by Clostridium perfringens epsilon toxin requires activation of P2 receptors

Large scale purification of Clostridium perfringens toxins: a review

Clostridium perfringens ε-Toxin Forms a Heptameric Pore within the Detergent-insoluble Microdomains of Madin-Darby Canine Kidney Cells and Rat Synaptosomes

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