Role of pannexin 1 in Clostridium perfringens beta-toxin-caused cell death.
Role of pannexin 1 in Clostridium perfringens beta-toxin-caused cell death.
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DOI：
10.1016/j.bbamem.2016.10.003
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发表时间：
2016-12
期刊：
影响因子：
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通讯作者：
Soshi Seike;Masaya Takehara;Keiko Kobayashi;M. Nagahama
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文献类型：
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作者：
Soshi Seike;Masaya Takehara;Keiko Kobayashi;M. Nagahama
产气荚膜梭菌产生的β-毒素是致死性出血性小肠结肠炎和肠毒血症的关键毒力因子。该毒素属于β-孔形成毒素（PFT）家族。我们最近报道了ATP门控的P2 X7受体与β-毒素相互作用。ATP释放通道泛连接蛋白1（Panx 1）是P2 X7受体信号转导的重要贡献者。因此，我们调查的参与Panx 1 β-毒素引起的细胞death.MethodsWe研究的影响Panx 1 β-毒素诱导的细胞death.MethodsWe利用选择性拮抗剂，敲低Panx 1，并结合斑点印迹分析。毒素处理后的Panx 1和P2 X7受体的定位测定免疫荧光staining.ResultsSelective Panx 1拮抗剂（甘珀酸[CBX]，丙磺舒，Panx 1抑制肽）防止β-毒素引起的细胞死亡THP-1细胞。CBX不阻断毒素与细胞的结合。Panx 1的小干扰敲低通过抑制毒素的寡聚体形成来阻断β毒素介导的细胞死亡。β-毒素触发THP-1细胞的瞬时ATP释放，但这种早期ATP释放被CBX阻断。ATP清除剂（腺苷三磷酸双磷酸酶和己糖激酶）抑制β-毒素诱导的细胞毒性。此外，共同管理的ATP与β-毒素增强的结合和细胞毒性的toxin.ConclusionsBased上我们的研究结果，Panx 1激活是通过β-毒素与P2 X7受体的相互作用。然后，由Panx 1通道开放释放的ATP促进毒素的寡聚体形成，导致细胞death.General SignificancePannexin 1是β-毒素介导的疾病的新的候选治疗靶点。
BackgroundBeta-toxin produced byClostridium perfringensis a key virulence factor of fatal hemorrhagic enterocolitis and enterotoxemia. This toxin belongs to a family of β-pore-forming toxins (PFTs). We reported recently that the ATP-gated P2X7receptor interacts with beta-toxin. The ATP-release channel pannexin 1 (Panx1) is an important contributor to P2X7receptor signaling. Hence, we investigated the involvement of Panx1 in beta-toxin-caused cell death.MethodsWe examined the effect of Panx1 in beta-toxin-induced cell death utilizing selective antagonists, knockdown of Panx1, and binding using dot-blot analysis. Localization of Panx1 and the P2X7receptor after toxin treatment was determined by immunofluorescence staining.ResultsSelective Panx1 antagonists (carbenoxolone [CBX], probenecid, and Panx1 inhibitory peptide) prevented beta-toxin-caused cell death in THP-1 cells. CBX did not block the binding of the toxin to cells. Small interfering knockdown of Panx1 blocked beta-toxin-mediated cell death through inhibiting the oligomer formation of the toxin. Beta-toxin triggered a transient ATP release from THP-1 cells, but this early ATP release was blocked by CBX. ATP scavengers (apyrase and hexokinase) inhibited beta-toxin-induced cytotoxicity. Furthermore, co-administration of ATP with beta-toxin enhanced the binding and cytotoxicity of the toxin.ConclusionsBased on our results, Panx1 activation is achieved through the interaction of beta-toxin with the P2X7receptor. Then, ATP released by the Panx1 channel opening promotes oligomer formation of the toxin, leading to cell death.General significancePannexin 1 is a novel candidate therapeutic target for beta-toxin-mediated disease.