Henry, RoselaniCobbold, SimonAllen, RichardKhan, AsifHayward, RhysLehane, AdeleBray, PatrickHowitt, SusanBiagini, Giancarlo ASaliba, KevinKirk, Kiaran2015-12-080021-9258http://hdl.handle.net/1885/34753The intraerythrocytic malaria parasite exerts tight control over its ionic composition. In this study, a combination of fluorescent ion indicators and36Cl- flux measurements was used to investigate the transport of Cl- and the Cl--dependent transport of "H+-equivalents" in mature (trophozoite stage) parasites, isolated from their host erythrocytes. Removal of extracellular Cl-, resulting in an outward [Cl-] gradient, gave rise to a cytosolic alkalinization (i.e. a net efflux of H+-equivalents). This was reversed on restoration of extracellular Cl-. The flux of H+-equivalents was inhibited by 4,4′-diisothiocyanostilbene-2, 2′-disulfonic acid and, when measured in ATP-depleted parasites, showed a pronounced dependence on the pH of the parasite cytosol; the flux was low at cytosolic pH values < 7.2 but increased steeply with cytosolic pH at values > 7.2.36Cl- influx measurements revealed the presence of a Cl- uptake mechanism with characteristics similar to those of the Cl--dependent H+-equivalent flux. The intracellular concentration of Cl- in the parasite was estimated to be ∼48 mM in situ. The data are consistent with the intraerythrocytic parasite having in its plasma membrane a 4,4′-diisothiocyanostilbene-2,2′-disulfonic acid-sensitive transporter that, under physiological conditions, imports Cl- together with H+-equivalents, resulting in an intracellular Cl- concentration well above that which would occur if Cl- ions were distributed passively in accordance with the parasite's large, inwardly negative membrane potential.Keywords: Cytosolic; Cytosols; Disulfonic acid; Extracellular; Flux measurements; In-situ; Intracellular concentration; Ionic composition; Malaria parasite; Membrane potentials; Parasite-; pH value; Physiological condition; Plasma membranes; Plasmodium falciparum;201010.1074/jbc.M110.1209802016-02-24