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J Biol Chem, Vol. 274, Issue 47, 33213-33219, November 19, 1999

pH Regulation in the Intracellular Malaria Parasite, Plasmodium falciparum
H⁺ EXTRUSION VIA A V-TYPE H⁺-ATPase

From the Division of Biochemistry and Molecular Biology, Faculty of Science, Australian National University, Canberra, Australian Capital Territory 0200, Australia

The mechanism by which the intra-erythrocytic form of the human malaria parasite, Plasmodium falciparum, extrudes H⁺ ions and thereby regulates its cytosolic pH (pH_i), was investigated using saponin-permeabilized parasitized erythrocytes. The parasite was able both to maintain its resting pH_i and to recover from an imposed intracellular acidification in the absence of extracellular Na⁺, thus ruling out the involvement of a Na⁺/H⁺ exchanger in both processes. Both phenomena were ATP-dependent. Amiloride and the related compound ethylisopropylamiloride caused a substantial reduction in the resting pH_i of the parasite, whereas EMD 96785, a potent and allegedly selective inhibitor of Na⁺/H⁺ exchange, had relatively little effect. The resting pH_i of the parasite was also reduced by the sulfhydryl reagent N-ethylmaleimide, by the carboxyl group blocker N,N'-dicyclohexylcarbodiimide, and by bafilomycin A₁, a potent inhibitor of V-type H⁺-ATPases. Bafilomycin A₁ blocked pH_i recovery in parasites subjected to an intracellular acidification and reduced the rate of acidification of a weakly buffered solution by parasites under resting conditions. The data are consistent with the hypothesis that the malaria parasite, like other parasitic protozoa, has in its plasma membrane a V-type H⁺-ATPase, which serves as the major route for the efflux of H⁺ ions.

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