Molecular cloning and functional characterization of the receptor for Clostridium perfringens enterotoxin.
Molecular cloning and functional characterization of the receptor for Clostridium perfringens enterotoxin.
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DOI：
10.1083/jcb.136.6.1239
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发表时间：
1997-03-24
期刊：
影响因子：
--
通讯作者：
Sugimoto N
中科院分区：
文献类型：
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作者：
Katahira J;Inoue N;Horiguchi Y;Matsuda M;Sugimoto N
从肠毒素敏感的Vero细胞表达文库中克隆了产气荚膜梭菌肠毒素受体基因(CPE-R)的编码基因。CPE-R的核苷酸序列分析表明，该肠毒素受体由209个氨基酸组成，计算相对分子质量为22,029 D。该受体高度疏水，含有4个可能的跨膜片段，与功能未知的大鼠雄激素戒断细胞凋亡蛋白RVP1和小鼠少突胶质细胞特异性蛋白RVP1有显著的相似性。CPE-R在肠毒素敏感的Vero、Hep3B和肠道407细胞中均有表达，而在肠毒素不敏感的K562和JY细胞中未见表达。在肠毒素抗性的L929细胞中表达的CPE-R基因产物与肠毒素特异性地、直接地、高亲和力地结合，使细胞对毒素敏感，表明克隆的受体具有功能。结果表明，肠毒素除非与受体相互作用，否则不能组装成具有一定结构的复合体。这些结果表明，肠毒素受体对靶细胞的识别和细胞膜上的后改造都是必需的。
A cDNA encoding the Clostridium perfringens enterotoxin receptor gene (CPE-R) was cloned from an expression library of enterotoxin-sensitive Vero cells. The nucleotide sequence of CPE-R showed that the enterotoxin receptor consists of 209 amino acids with a calculated molecular mass of 22,029 D. This receptor is highly hydrophobic, contains four putative transmembrane segments, and has significant similarity to the rat androgen withdrawal apoptosis protein RVP1 and the mouse oligodendrocyte specific protein, the functions of which are unknown. The expression of CPE-R was detected in the enterotoxin-sensitive Vero, Hep3B, and Intestine 407 cell lines, but not in the enterotoxin-insensitive K562 and JY cell lines. The CPE-R gene product expressed in enterotoxin-resistant L929 cells bound to enterotoxin specifically and directly and with high affinity and rendered the cells sensitive to the toxin, indicating that the cloned receptor is functional. Results showed that enterotoxin could not assemble into a complex with a defined structure unless it interacted with the receptor. From these results, it is proposed that the enterotoxin receptor is required for both target cell recognition and poreformation in the cell membrane.