Clostridium perfringens enterotoxin utilizes two structurally related membrane proteins as functional receptors in vivo.
Human and mouse cDNAs showing homology to the Clostridium perfringens enterotoxin (CPE) receptor gene (CPE-R) from Vero cells (DDBJ/EMBL/GenBankTM accession no. D88492) (Katahira, J., Inoue, N., Horiguchi, Y., Matsuda, M., and Sugimoto, N. (1997) J. Cell Biol. 136, 1239-1247) were cloned. They were classified into two groups, the Vero cell CPE receptor homologues and rat androgen withdrawal apoptosis protein (RVP1; accession no. M74067) homologues, based on the similarities of primary amino acid sequences. L929 cells that were originally insensitive to CPE became sensitive to CPE on their transfection with cDNAs encoding either the CPE receptor or RVP1 homologues, indicating that these gene products are not only structurally similar but also functionally active as receptors for CPE. By binding assay, the human RVP1 homologue showed differences in affinity and capacity of binding from those of the human CPE receptor. Northern blot analysis showed that mouse homologues of the CPE receptor and RVP1 are expressed abundantly in mouse small intestine. The expression of CPE-R mRNA in the small intestine was restricted to cryptic enterocytes, indicating that the CPE receptor is expressed in intestinal epithelial cells. These results are consistent with reports that CPE binds to the small intestinal cells via two different kinds of receptors. High levels of expression of CPE-R and/or RVP1 mRNA were also detected in other organs, including the lungs, liver, and kidneys, but only low levels were expressed in heart and skeletal muscles. These results indicate that CPE uses structurally related cellular proteins as functional receptors in vivo and that organs that have not so far been recognized as CPE-sensitive have the potential to be targets of CPE.
Katahira
J
J
Department of Bacterial Toxinology, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamadaoka, Suita, Osaka 565, Japan. c72@ix.urz.uni-heidelberg.de
Sugiyama
H
H
Inoue
N
N
Horiguchi
Y
Y
Matsuda
M
M
Sugimoto
N
N
D000595:Amino Acid Sequence; D000818:Animals; D002522:Chlorocebus aethiops; D062465:Claudin-3; D062506:Claudin-4; D003016:Clostridium perfringens / Q000378:metabolism*; D018076:DNA, Complementary; D004768:Enterotoxins / Q000378:metabolism*; D006801:Humans; D007421:Intestine, Small / Q000378:metabolism; D007700:Kinetics; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000737:chemistry / Q000378:metabolism*; D014709:Vero Cells
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; D008565:Membrane Proteins; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; C035598:enterotoxin, Clostridium; C105422:Clostridium perfringens enterotoxin receptor