Identification and characterization of phospholipid metabolic enzymes in Giardia lamblia

TL;DRAbstract

As part of our efforts in understanding the mechanism of lipid scavenging and remodeling reactions in Giardia lamblia trophozoites, we have analyzed the location and function of specific phospholipid metabolic enzymes. AU1 epitope tagged constructs were generated for a lysophospholipase enzyme (GLPL) gene and individually transfected into Giardia trophozoites. Confocal analysis revealed that the expression of the AU1 epitope tagged GLPL concentrated in endoplasmic reticulum/perinuclear regions and co-localized with labeled ceramide and sphingomyelin. This is to be expected for a phospholipase A2 enzyme. To test this hypothesis, we generated giardiavirus Hammerhead ribozyme constructs to knock down mRNA for GLPL as well as a second identified enzyme in the lipid-remodeling pathway, lysophosphatydic acid acyl transferase (GLAAT). Giardia trophozoites transfected with either knockout construct demonstrated an initial lower total cell number compared to sham constructs, producing irregular

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