Neutral sphingomyelinase activity dependent on Mg2+ and anionic phospholipids in the intraerythrocytic malaria parasite Plasmodium falciparum.
Sphingolipid metabolism and metabolites are important in various cellular events in eukaryotes. However, little is known about their function in plasmodial parasites. Here we demonstrate that neutral sphingomyelinase (SMase) involved in the sphingomyelin (SM) catabolism is retained by the intraerythrocytic parasite Plasmodium falciparum. When assayed in a neutral pH buffer supplemented with Mg(2+) and phosphatidylserine, an activity for the release of the phosphocholine group from SM was detected in parasite-infected, but not in uninfected, erythrocyte ghosts. The SMase activity in the parasite-infected erythrocyte ghosts was enhanced markedly by anionic phospholipids including unsaturated but not saturated phosphatidylserine. Mn(2+) could not substitute for Mg(2+) to activate SMase in parasite-infected erythrocyte ghosts, whereas both Mn(2+) and Mg(2+) activated mammalian neutral SMase. The specific activity level of SMase was higher in isolated parasites than in infected erythrocyte ghosts; further fractionation of lysates of the isolated parasites showed that the activity was bound largely to the membrane fraction of the parasites. The plasmodial SMase seemed not to hydrolyse phosphatidylcholine or phosphatidylinositol. The plasmodial SMase, but not SM synthase, was sensitive to scyphostatin, an inhibitor of mammalian neutral SMase, indicating that the plasmodial activities for SM hydrolysis and SM synthesis are mediated by different catalysts. Our finding that the malaria parasites possess SMase activity might explain why the parasites seem to have an SM synthase activity but no activity to synthesize ceramide de novo.
Hanada
K
K
Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, 1-23-1, Toyama, Shinjuku-ku, Tokyo 162-8640, Japan. hanak@nih.go.jp
Mitamura
T
T
Fukasawa
M
M
Magistrado
P A
PA
Horii
T
T
Nishijima
M
M
D000577:Amides / Q000494:pharmacology; D000818:Animals; D000838:Anions; D002413:Cations, Divalent; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004912:Erythrocytes / Q000469:parasitology; D006801:Humans; D008274:Magnesium / Q000378:metabolism*; D008614:Meperidine / Q000031:analogs & derivatives / Q000494:pharmacology; D010743:Phospholipids / Q000378:metabolism*; D010963:Plasmodium falciparum / Q000201:enzymology / Q000378:metabolism*; D011753:Pyrones / Q000494:pharmacology; D013108:Sphingomyelin Phosphodiesterase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D013379:Substrate Specificity
D000577:Amides; D000838:Anions; D002413:Cations, Divalent; D004791:Enzyme Inhibitors; D010743:Phospholipids; D011753:Pyrones; C121123:scyphostatin; C026733:4-propionyloxy-4-phenyl-N-methylpiperidine; D008614:Meperidine; D013108:Sphingomyelin Phosphodiesterase; D008274:Magnesium