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Fig. 1. Alkalinisation of the DV following inhibition of the V-type H+-ATPase by concanamycin A. (A,B) Representative fluorometer traces for mature trophozoite-stage D10 (CQS; A) and 7G8 (CQR; B) parasites. The parasites were isolated using saponin-permeabilisation of the erythrocyte and parasitophorous vacuole membranes, and suspended in minimal saline solution at pH 7.1. The H+ ionophore CCCP (100 nM) or solvent (DMSO) was added to parasites 1 minute prior to the addition of concanamycin A (100 nM). In A and B the baseline fluorescence ratio (before the addition of concanamycin A) is that for the DMSO control trace. Concanamycin A addition is indicated by black arrowheads. Traces are representative of those obtained in seven independent experiments for 7G8 parasites and 12 independent experiments for D10 parasites. (C) Representative fluorometer trace for saponin-isolated mature-trophozoite-stage D10 parasites to which 10 µM CCCP was added (indicated by white arrowhead) 1 minute prior to the addition of 100 nM concanamycin A (black arrowhead). Similar data were obtained with other strains (not shown). (D) Averaged data for the rate of DV alkalinisation (expressed as the inverse of the alkalinisation half-time) in the CQS strains D10 and 3D7 and in the CQR strains 7G8 and K1, to which had been added (1 minute before the addition of 100 nM concanamycin A) either CCCP in DMSO (giving a final CCCP concentration of 100 nM) or DMSO alone. Data are the average from at least five independent experiments (+ s.e.m.) for each condition and strain.
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