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.
Highlights
Malaria is one of the most lethal parasitic human diseases in the developing world, causing about half a million deaths annually [1]
In order to assess whether the P. falciparum E-NTPDase is important for parasite development, we incubated P. falciparum with the ENTPDase inhibitors suramin, ARL 67156, and gadolinium for 48 h and measured the parasitemia at different time points by flow cytometry (Fig. 2)
Gadolinium was capable of inhibiting the E-NTPDase from Torpedo electric organ and ecto-ATPase of T. cruzi [16, Table 1 Primer sequences for the quantitative real-time PCR (qRT-PCR) analysis and cloning of the green fluorescent protein (GFP)-fusion construct
Summary
Malaria is one of the most lethal parasitic human diseases in the developing world, causing about half a million deaths annually [1]. Its etiological agent belongs to the genus Plasmodium, and among these, Plasmodium falciparum is the one responsible for the most severe form of the disease [2]. There are reported cases of parasite resistance to all available anti-malarial drugs, and the understanding of the parasite physiology and signaling events will help to identify new drugs targets [4,5,6]. Components of the signaling machinery are being considered potential drug targets in malaria parasites. It is well known that Plasmodium is able to convert external stimuli into intracellular responses, [7,8,9,10]
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
