Molecular cloning and functional characterization of the receptor for Clostridium perfringens enterotoxin.
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 pore formation in the cell membrane.
Katahira
J
J
Department of Bacterial Toxicology, Osaka University, Japan. katahira@biken.osaka-u.ac.jp
Inoue
N
N
Horiguchi
Y
Y
Matsuda
M
M
Sugimoto
N
N
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line / Q000187:drug effects; D002522:Chlorocebus aethiops / Q000235:genetics; D062465:Claudin-3; D062506:Claudin-4; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004351:Drug Resistance; D004768:Enterotoxins / Q000378:metabolism / Q000494:pharmacology*; D005434:Flow Cytometry; D015723:Gene Library; D005796:Genes; D006801:Humans; D007739:L Cells; D046911:Macromolecular Substances; D008565:Membrane Proteins; D051379:Mice; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000378:metabolism; D011506:Proteins / Q000737:chemistry; D051381:Rats; D011956:Receptors, Cell Surface / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D014407:Tumor Cells, Cultured / Q000187:drug effects; D014709:Vero Cells / Q000737:chemistry
C568122:CLDN3 protein, human; C568157:CLDN4 protein, human; D062465:Claudin-3; D062506:Claudin-4; C568123:Cldn3 protein, mouse; C568124:Cldn3 protein, rat; C568158:Cldn4 protein, mouse; D018076:DNA, Complementary; D004768:Enterotoxins; D046911:Macromolecular Substances; D008565:Membrane Proteins; D010446:Peptide Fragments; D011506:Proteins; D011956:Receptors, Cell Surface; C035598:enterotoxin, Clostridium; C105422:Clostridium perfringens enterotoxin receptor