| Article ID | Journal | Published Year | Pages | File Type |
|---|---|---|---|---|
| 6292279 | Experimental Parasitology | 2010 | 7 Pages |
Abstract
A novel neutral sphingomyelinase (nSMase) was characterized in Entamoeba histolytica trophozoites. SMase, a sphingomyelin-specific form of phospholipase C, catalyzes the hydrolysis of sphingomyelin to ceramide and phosphorylcholine. Three amebic putative nSMase genes were found to be actively transcribed. Mg2+-independent nSMase activity in the soluble fraction of the trophozoites was stimulated by Mn2+ and partially inhibited by Zn2+. nSMase activity of the recombinant protein EhnSM1, increased 4.5-fold in the presence of 0.5Â mM Mn2+, and abolished by 5Â mM Zn2+. A dose-dependent inhibition of rEhnSM1 was observed with scyphostatin, a specific inhibitor of nSMases. The EhnSM1 and EhnSM3 were detected in the soluble fraction of the amebic lysate as 35-37Â kDa proteins by western blot analysis. Immunofluorescence assay showed that the overexpressed HA-tagged EhnSM1 and EhnSM3 were localized to the cytosol. The biological role of these novel E. histolytica nSMases described in this work remains to be determined.
Keywords
PBSnSMaseRT-PCRRNSIFASMaseASMaseEntamoeba histolyticaROSimmunofluorescence assayacidic sphingomyelinasesphingomyelinsphingomyelinaseneutral sphingomyelinaseceramidePhosphatidylserinePhosphate-buffered salinehemagglutininreverse transcriptase polymerase chain reactionreactive nitrogen speciesReactive oxygen species
Related Topics
Life Sciences
Immunology and Microbiology
Parasitology
Authors
Claudia Leticia Mendoza-MacÃas, Minerva Paola Barrios-Ceballos, Fernando Anaya-Velázquez, Kumiko Nakada-Tsukui, Tomoyoshi Nozaki, Felipe Padilla-Vaca,