A plant-like vacuolar H(+)-pyrophosphatase in Plasmodium falciparum.

Inorganic pyrophosphate promoted the acidification of a subcellular compartment in cell homogenates of Plasmodium falciparum trophozoites. The proton gradient driven by pyrophosphate was collapsed by addition of NH(4)Cl or the K(+)/H(+) exchanger nigericin and eliminated by the pyrophosphate analog aminomethylenediphosphonate. Pyrophosphatase activity was dependent upon K(+), and partially inhibited by Na(+). The presence of a plant-like vacuolar H(+)-translocating pyrophosphatase (V-H(+)-PPase) was confirmed using antibodies raised against conserved peptide sequences of the enzyme, which cross reacted with a protein band of 76.5 kDa. Immunofluorescence microscopy using these antibodies showed a general fluorescence over the whole parasites and intracellular bright spots suggesting a vesicular and plasma membrane localization. Together, these results indicate the presence in P. falciparum of a V-H(+)-PPase of similar characteristics to those of the enzyme from plants.