title: Clostridium perfringens alpha-toxin activates the sphingomyelin metabolism system in sheep erythrocytes.
abstract: Clostridium perfringens alpha-toxin induces hemolysis of rabbit erythrocytes through the activation of glycerophospholipid metabolism. Sheep erythrocytes contain large amounts of sphingomyelin (SM) but not phosphatidylcholine. We investigated the relationship between the toxin-induced hemolysis and SM metabolic system in sheep erythrocytes. Alpha-toxin simultaneously induced hemolysis and a reduction in the levels of SM and formation of ceramide and sphingosine 1-phosphate (S1P). N-Oleoylethanolamine, a ceramidase inhibitor, inhibited the toxin-induced hemolysis and caused ceramide to accumulate in the toxin-treated cells. Furthermore, dl-threo-dihydrosphingosine and B-5354c, isolated from a novel marine bacterium, both sphingosine kinase inhibitors, blocked the toxin-induced hemolysis and production of S1P and caused sphingosine to accumulate. These observations suggest that the toxin-induced activation of the SM metabolic system is closely related to hemolysis. S1P potentiated the toxin-induced hemolysis of saponin-permeabilized erythrocytes but had no effect on that of intact cells. Preincubation of lysated sheep erythrocytes with pertussis toxin blocked the alpha-toxin-induced formation of ceramide from SM. In addition, incubation of C. botulinum C3 exoenzyme-treated lysates of sheep erythrocytes with alpha-toxin caused an accumulation of sphingosine and inhibition of the formation of S1P. These observations suggest that the alpha-toxin-induced hemolysis of sheep erythrocytes is dependent on the activation of the SM metabolic system through GTP-binding proteins, especially the formation of S1P.
authors.affiliation: Department of Microbiology, Fujita Health University, School of Medicine, Toyoake, Aichi 470-1192, Japan.
authors.forename: Sadayuki
authors.initials: S
authors.lastname: Ochi
authors.forename: Masataka
authors.initials: M
authors.lastname: Oda
authors.forename: Hisaaki
authors.initials: H
authors.lastname: Matsuda
authors.forename: Syusuke
authors.initials: S
authors.lastname: Ikari
authors.forename: Jun
authors.initials: J
authors.lastname: Sakurai
mesh_terms: D010129:4-Aminobenzoic Acid / Q000494:pharmacology; D036002:ADP Ribose Transferases / Q000494:pharmacology; D000581:Amidohydrolases / Q000037:antagonists & inhibitors; D000818:Animals; D001427:Bacterial Toxins / Q000378:metabolism*; D001905:Botulinum Toxins / Q000494:pharmacology; D002135:Calcium-Binding Proteins; D055572:Ceramidases; D002855:Chromatography, Thin Layer; D004075:Diglycerides / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D063388:Endocannabinoids; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004912:Erythrocytes / Q000378:metabolism*; D004983:Ethanolamines / Q000737:chemistry / Q000494:pharmacology; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000378:metabolism; D006461:Hemolysis; D015544:Inositol 1,4,5-Trisphosphate / Q000494:pharmacology; D008246:Lysophospholipids / Q000737:chemistry; D009829:Oleic Acids; D037342:Pertussis Toxin / Q000494:pharmacology; D010713:Phosphatidylcholines / Q000494:pharmacology; D010767:Phosphorylcholine / Q000378:metabolism; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000378:metabolism; D011817:Rabbits; D012756:Sheep; D013109:Sphingomyelins / Q000378:metabolism*; D013110:Sphingosine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D013997:Time Factors; D014118:Toxins, Biological; D010738:Type C Phospholipases / Q000378:metabolism*; D062366:para-Aminobenzoates; D000099157:Sphingosine Kinase
chemical_list: D010129:4-Aminobenzoic Acid; D036002:ADP Ribose Transferases; D000581:Amidohydrolases; D001427:Bacterial Toxins; D001905:Botulinum Toxins; D002135:Calcium-Binding Proteins; D055572:Ceramidases; D004075:Diglycerides; D063388:Endocannabinoids; D004791:Enzyme Inhibitors; D004983:Ethanolamines; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D006160:Guanosine Triphosphate; D015544:Inositol 1,4,5-Trisphosphate; D008246:Lysophospholipids; D009829:Oleic Acids; D037342:Pertussis Toxin; D010713:Phosphatidylcholines; D010767:Phosphorylcholine; D017853:Phosphotransferases (Alcohol Group Acceptor); D013109:Sphingomyelins; D013110:Sphingosine; D014118:Toxins, Biological; D010738:Type C Phospholipases; D062366:para-Aminobenzoates; D000099157:Sphingosine Kinase; C011439:1,2-diacylglycerol; C416812:B 5354c; C033595:N-oleoylethanolamine; C060506:sphingosine 1-phosphate; C063007:exoenzyme C3, Clostridium botulinum; C040649:alpha toxin, Clostridium perfringens; C005682:safingol