title: Transport of nucleosides across the Plasmodium falciparum parasite plasma membrane has characteristics of PfENT1.
abstract: Like all parasitic protozoa, the human malaria parasite Plasmodium falciparum lacks the enzymes required for de novo synthesis of purines and it is therefore reliant upon the salvage of these compounds from the external environment. P. falciparum equilibrative nucleoside transporter 1 (PfENT1) is a nucleoside transporter that has been localized to the plasma membrane of the intraerythrocytic form of the parasite. In this study we have characterized the transport of purine and pyrimidine nucleosides across the plasma membrane of 'isolated' trophozoite-stage P. falciparum parasites and compared the transport characteristics of the parasite with those of PfENT1 expressed in Xenopus oocytes. The transport of nucleosides into the parasite: (i) was, in the case of adenosine, inosine and thymidine, very fast, equilibrating within a few seconds; (ii) was of low affinity [K(m) (adenosine) = 1.45 +/- 0.25 mM; K(m) (thymidine) = 1.11 +/- 0.09 mM]; and (iii) showed 'cross-competition' for adenosine, inosine and thymidine, but not cytidine. The kinetic characteristics of nucleoside transport in intact parasites matched very closely those of PfENT1 expressed in Xenopus oocytes [K(m) (adenosine) = 1.86 +/- 0.28 mM; K(m) (thymidine) = 1.33 +/- 0.17 mM]. Furthermore, PfENT1 transported adenosine, inosine and thymidine, with a cross-competition profile the same as that seen for isolated parasites. The data are consistent with PfENT1 serving as a major route for the uptake of nucleosides across the parasite plasma membrane.
authors.affiliation: School of Biochemistry and Molecular Biology, The Australian National University, Canberra ACT 0200, Australia.
authors.forename: Megan J
authors.initials: MJ
authors.lastname: Downie
authors.forename: Kevin J
authors.initials: KJ
authors.lastname: Saliba
authors.forename: Susan M
authors.initials: SM
authors.lastname: Howitt
authors.forename: Stefan
authors.initials: S
authors.lastname: Bröer
authors.forename: Kiaran
authors.initials: K
authors.lastname: Kirk
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000378:metabolism*; D004912:Erythrocytes / Q000469:parasitology; D006801:Humans; D033701:Nucleobase, Nucleoside, Nucleotide, and Nucleic Acid Transport Proteins / Q000378:metabolism*; D033703:Nucleoside Transport Proteins / Q000378:metabolism; D009865:Oocytes / Q000378:metabolism; D010963:Plasmodium falciparum / Q000254:growth & development / Q000378:metabolism*; D015800:Protozoan Proteins / Q000378:metabolism*; D011687:Purines / Q000378:metabolism*; D011743:Pyrimidines / Q000378:metabolism*; D014981:Xenopus / Q000254:growth & development / Q000378:metabolism
chemical_list: C430711:ENT1 protein, Plasmodium falciparum; D033701:Nucleobase, Nucleoside, Nucleotide, and Nucleic Acid Transport Proteins; D033703:Nucleoside Transport Proteins; D015800:Protozoan Proteins; D011687:Purines; D011743:Pyrimidines