<Emphasis Type="Italic">Plasmodium</Emphasis><Emphasis Type="Italic">falciparum</Emphasis> GFP-E-NTPDase expression at the intraerythrocytic stages and its inhibition blocks the development of the human malaria parasite |
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Authors: | Lucas Borges-Pereira Kamila Anna Meissner Carsten Wrenger Célia R S Garcia |
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Institution: | 1.Departamento de Parasitologia, Instituto de Ciências Biomédicas,Universidade de S?o Paulo,S?o Paulo,Brazil;2.Departamento de Fisiologia, Instituto de Biociências,Universidade de S?o Paulo,S?o Paulo,Brazil |
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Abstract: | Plasmodium falciparum is the causative agent of the most dangerous form of malaria in humans. It has been reported that the P. falciparum genome encodes for a single ecto-nucleoside triphosphate diphosphohydrolase (E-NTPDase), an enzyme that hydrolyzes extracellular tri- and di-phosphate nucleotides. The E-NTPDases are known for participating in invasion and as a virulence factor in many pathogenic protozoa. Despite its presence in the parasite genome, currently, no information exists about the activity of this predicted protein. Here, we show for the first time that P. falciparum E-NTPDase is relevant for parasite lifecycle as inhibition of this enzyme impairs the development of P. falciparum within red blood cells (RBCs). ATPase activity could be detected in rings, trophozoites, and schizonts, as well as qRT-PCR, confirming that E-NTPDase is expressed throughout the intraerythrocytic cycle. In addition, transfection of a construct which expresses approximately the first 500 bp of an E-NTPDase-GFP chimera shows that E-NTPDase co-localizes with the endoplasmic reticulum (ER) in the early stages and with the digestive vacuole (DV) in the late stages of P. falciparum intraerythrocytic cycle. |
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