Cholesterol and lipid/protein ratio control the oligomerization of a sphingomyelin-specific toxin, lysenin.
Lysenin is a pore-forming toxin that specifically binds sphingomyelin (SM). The binding of the toxin to the membrane is accompanied by the oligomerization of the protein, leading to pore formation. The interaction of lysenin with SM is affected by the presence of other lipids found in the plasma membrane. Although a previous study showed that SM/cholesterol liposomes were 10,000 times more effective than SM liposomes in inhibiting lysenin-induced hemolysis (Yamaji, A., Sekizawa, Y., Emoto, K., Sakuraba, H., Inoue, K., Kobayashi, H., and Umeda, M. (1998) J. Biol. Chem. 273, 5300-5306), the role of cholesterol is not precisely clarified. In the present study, we examined the effects of the presence of cholesterol in the SM membrane on the inhibition of hemolysis, the binding of lysenin to SM, and the oligomerization of lysenin. The addition of cholesterol to SM liposomes dramatically inhibited lysenin-induced hemolysis as described previously. However, the presence of cholesterol did not affect the binding of lysenin to SM liposomes. The oligomerization of lysenin was facilitated by the presence of cholesterol in SM liposomes. The oligomerization of lysenin was also dependent on the SM/lysenin ratio, that is, the amount of lysenin oligomer was increased with the decrease in the SM/lysenin ratio. When the SM/lysenin molar ratio was high, lysenin associated with the membrane as a monomer, which was able to transfer to the erythrocyte membrane. Our results indicate that both cholesterol and the SM/lysenin ratio control the amount of lysenin monomer via altering the state of protein oligomerization, thus affecting hemolysis.
Ishitsuka
Reiko
R
Lipid Biology Laboratory, RIKEN (Institute of Physical and Chemical Research) Discovery Research Institute, 2-1 Hirosawa, Wako-shi, Saitama 351-0198, Japan.
Kobayashi
Toshihide
T
D000818:Animals; D002784:Cholesterol / Q000737:chemistry / Q000494:pharmacology*; D004912:Erythrocytes / Q000187:drug effects; D006461:Hemolysis / Q000187:drug effects; D008081:Liposomes / Q000737:chemistry; D009835:Oligochaeta / Q000737:chemistry; D010713:Phosphatidylcholines / Q000737:chemistry; D017434:Protein Structure, Tertiary; D012756:Sheep; D013109:Sphingomyelins / Q000737:chemistry*; D014118:Toxins, Biological / Q000737:chemistry* / Q000494:pharmacology
D008081:Liposomes; D010713:Phosphatidylcholines; D013109:Sphingomyelins; D014118:Toxins, Biological; C107040:lysenin; D002784:Cholesterol; C017251:1,2-oleoylphosphatidylcholine