Compositional and stoichiometric analysis of Clostridium perfringens enterotoxin complexes in Caco-2 cells and claudin 4 fibroblast transfectants.
Compositional and stoichiometric analysis of Clostridium perfringens enterotoxin complexes in Caco-2 cells and claudin 4 fibroblast transfectants.
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Caco-2 细胞和 Claudin 4 成纤维细胞转染子中产气荚膜梭菌肠毒素复合物的组成和化学计量分析。
DOI：
10.1111/j.1462-5822.2007.00994.x
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发表时间：
2007
影响因子：
3.4
通讯作者：
McClane,BruceA
中科院分区：
文献类型：
--
作者：
Robertson,SusanL;Smedley3rd,JamesG;Singh,Usha;Chakrabarti,Ganes;VanItallie,ChristinaM;Anderson,JamesM;McClane,BruceA
Clostridium perfringensenterotoxin (CPE) binds to host cell receptors, forming a small complex precursor for two large complexes reportedly having molecular masses of ∼155 or ∼200 kDa. Formation of the ∼155 kDa complex causes a Ca2+influx that leads to apoptosis or oncosis. CPE complex composition is currently poorly understood, although occludin was identified in the ∼200 kDa complex. The current study used heteromer gel shift analysis to show both CPE large complexes contain six CPE molecules. Ferguson plots and size exclusion chromatography re‐sized the ∼155 and ∼200 kDa complexes as ∼425–500 kDa and ∼550–660 kDa respectively. Co‐immunoprecipitation and electroelution studies demonstrated both CPE‐binding and non‐CPE‐binding claudins are associated with all three CPE complexes in Caco‐2 cells and with small complex and ∼425–500 kDa complex of claudin 4 transfectants. Fibroblast transfectants expressing claudin 4 or C‐terminal truncated claudin 4 were CPE‐sensitive and formed the ∼425 kDa complex, indicating claudin‐induced cell signalling is not required for CPE action and that expression of a single receptor claudin suffices for ∼425–500 kDa CPE complex formation. These results identify CPE as a unique toxin that combines with tight junction proteins to form high‐molecular‐mass hexameric pores and alter membrane permeability.
