Mg(2+)-ATP-dependent H(+)-translocation has been studied in membrane vesicles derived from the roots of Gossypium hirsutum L. var. Acala San Jose 2. Establishment of a positive membrane potential was followed by measuring SCN(-) accumulation; establishment of DeltapH across the vesicle membranes by measuring quinacrine fluorescence quenching. High specificity for ATP was shown, and H(+)-translocation was oligomycin stable. The pH profile for H(+)-translocation showed an optimum at 5.5. The relationship between SCN(-) accumulation and ATP concentration was approximately Michaelian; the apparent K(m) was 0.7 millimolar. K-2-(N-morpholino)ethanesulfonic acid strongly promoted ATP-dependent SCN(-) uptake (up to 180% stimulation). The effect was not given by Na-Mes. Carbonyl cyanide p-trifluoromethoxyphenylhydrazone totally inhibited SCN(-) accumulation, both in the presence and absence of K-2(N-morpholino)ethanesulfonic acid. Vanadate at 200 micromolar inhibited SCN(-) uptake by about 10 to 40% in the absence of K(+), but more strongly in its presence (about 60%). NO(3) (-) at 100 millimolar inhibited initial rate of quinacrine quenching by about 25%. The NO(3) (-) insensitive fraction was activated by K(+); and inhibited by 200 micromolar vanadate to about 40%, provided K(+) was present. Saline conditions during the growth of the plants had no appreciable effect on the observed characteristics of H(+)-translocation.
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Hassidim et al. (1986) studied this question.