Evidence for an Amiloride-Inhibited Mg2+/2H+ Antiporter in Lutoid (Vacuolar) Vesicles from Latex of Hevea brasiliensis

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Abstract
Lutoids represent a lysosomal microvacuolar compartment of rubber-tree (Hevea brasiliensis) latex. We observed acidification of isolated vesicles after imposing an outward Mg2+ diffusion gradient and dissipation of a preformed pH gradient in the presence of exogenous Mg2+. These results suggest the presence of a Mg2+/H+ antiporter. The maximum Mg2+/H+ exchange rate was observed at pH 8.5. The Km values for Mg2+ (2.6 mm) were identical for both influx and efflux experiments. When membrane potential was clamped at zero with K+ and valinomycin, the response of the membrane potential probe oxonol VI showed that the Mg2+/H+ exchange was electroneutral. Mg2+/H+ exchange was inhibited by amiloride and imipramine. Both the inhibiting concentration range and the Km for Mg2+ are similar to those reported for the Mg2+/2Na+ antiporter in animals cell. These data are consistent with the existence of a Mg2+/2H+ antiporter in a plant tonoplast.