title: Toxoplasma gondii tachyzoites possess an unusual plasma membrane adenosine transporter.
abstract: Nucleoside transport may play a critical role in successful intracellular parasitism by Toxoplasma gondii. This protozoan is incapable of de novo purine synthesis, and must salvage purines from the host cell. We characterized purine transport by extracellular T. gondii tachyzoites, focusing on adenosine, the preferred salvage substrate. Although wild-type RH tachyzoites concentrated [3H]adenosine 1.8-fold within 30 s, approx. half of the [3H]adenosine was converted to nucleotide, consistent with the known high parasite adenosine kinase activity. Studies using an adenosine kinase deficient mutant confirmed that adenosine transport was non-concentrative. [14C]Inosine, [14C]hypoxanthine and [3H]adenine transport was also rapid and non-concentrative. Adenosine transport was inhibited by dipyridamole (IC50 approx. 0.7 microM), but not nitrobenzylthioinosine (15 microM). Transport of inosine, hypoxanthine and adenine was minimally inhibited by 10 microM dipyridamole, however. Competition experiments using unlabeled nucleosides and bases demonstrated distinct inhibitor profiles for [3H]adenosine and [14C]inosine transport. These results are most consistent with a single, dipyridamole-sensitive, adenosine transporter located in the T. gondii plasma membrane. Additional permeation pathways for inosine, hypoxanthine, adenine and other purines may also be present.
authors.affiliation: Department of Internal Medicine, Yale University School of Medicine, New Haven, CT 06520-8022, USA.
authors.forename: J C
authors.initials: JC
authors.lastname: Schwab
authors.forename: M
authors.initials: M
authors.lastname: Afifi Afifi
authors.forename: G
authors.initials: G
authors.lastname: Pizzorno
authors.forename: R E
authors.initials: RE
authors.lastname: Handschumacher
authors.forename: K A
authors.initials: KA
authors.lastname: Joiner
mesh_terms: D000241:Adenosine / Q000378:metabolism*; D000248:Adenosine Kinase / Q000235:genetics / Q000378:metabolism; D000818:Animals; D001667:Binding, Competitive; D001692:Biological Transport; D002352:Carrier Proteins / Q000187:drug effects / Q000378:metabolism*; D004176:Dipyridamole / Q000494:pharmacology; D008565:Membrane Proteins / Q000187:drug effects / Q000378:metabolism*; D033703:Nucleoside Transport Proteins; D015800:Protozoan Proteins / Q000187:drug effects / Q000378:metabolism*; D011687:Purines / Q000378:metabolism; D013868:Thioinosine / Q000031:analogs & derivatives / Q000494:pharmacology; D014122:Toxoplasma / Q000737:chemistry* / Q000254:growth & development
chemical_list: D002352:Carrier Proteins; D008565:Membrane Proteins; D033703:Nucleoside Transport Proteins; D015800:Protozoan Proteins; D011687:Purines; D013868:Thioinosine; D004176:Dipyridamole; D000248:Adenosine Kinase; C001789:4-nitrobenzylthioinosine; D000241:Adenosine