The relationship between the metabolism of sphingomyelin species and the hemolysis of sheep erythrocytes induced by Clostridium perfringens alpha-toxin.

Abstract

Clostridium perfringens alpha-toxin induces the hemolysis of sheep erythrocytes by activating the metabolism of sphingomyelin (SM) via a GTP binding protein in membranes. alpha-Toxin stimulated the formation of 15-N-nervonoyl sphingosine (C24:1-ceramide), which was identified by positive ion fast atom bombardment-MS and 1H-NMR spectroscopy. C24:1-ceramide stimulated the toxin-induced hemolysis of saponin-pretreated sheep erythrocytes and increased the production of sphingosine 1-phosphate (S1P) in the cells, but N-lignoceroyl sphingosine did not. These events elicited by the toxin in the presence of C24:1-ceramide were significantly attenuated by treatment with dihydrosphingosine, a sphingosine kinase inhibitor. TLC showed that the level of C24:1-ceramide was highest among the ceramides with an unsaturated bond in the fatty acyl chain in the detergent-resistant membranes (DRMs). The toxin specifically bound to DRMs rich in cholesterol, resulting in the hydrolysis of N-nervonoic sphingomyelin (C24:1-SM) in DRMs. Treatment of the cells with pertussis toxin (PT) inhibited the alpha-toxin-induced formation of C24:1-ceramide from C24:1-SM in DRMs and hemolysis, indicating that endogenous sphingomyelinase, which hydrolyzes C24:1-SM to C24:1-ceramide, is controlled by PT-sensitive GTP binding protein in membranes. These results show that the toxin-induced metabolism of C24:1-SM to S1P in DRMs plays an important role in the toxin-induced hemolysis of sheep erythrocytes.

• Authors: Masataka Oda, Takayuki Matsuno, Ryouta Shiihara, Sadayuki Ochi, Rieko Yamauchi, Yuki Saito, Hiroshi Imagawa, Masahiro Nagahama, Mugio Nishizawa, Jun Sakurai
• Journal: Journal of lipid research (J Lipid Res) Vol. 49 Issue 5 Pg. 1039-47 (May 2008) ISSN: 0022-2275 [Print] United States
• PMID: 18263851 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Bacterial Toxins
• Calcium-Binding Proteins
• Detergents
• Sphingomyelins
• Type C Phospholipases
• alpha toxin, Clostridium perfringens
• Sphingosine

Topics

• Animals
• Bacterial Toxins (toxicity)
• Calcium-Binding Proteins (toxicity)
• Detergents (pharmacology)
• Erythrocyte Membrane (drug effects, physiology)
• Erythrocytes (drug effects, physiology)
• Hemolysis
• Kinetics
• Sheep
• Sphingomyelins (metabolism)
• Sphingosine (analogs & derivatives, pharmacology)
• Type C Phospholipases (toxicity)