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Plant Physiology 78:163-167 (1985)
© 1985 American Society of Plant Biologists

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Articles

Na+/H+ Antiport in Isolated Tonoplast Vesicles from Storage Tissue of Beta vulgaris1

Eduardo Blumwald and Ronald J. Poole

Department of Biology, McGill University, 1205 Avenue Docteur Penfield, Montreal, Quebec, Canada H3A 1B1

The pH-dependent fluorescence quenching of acridine orange was used to study the Na+- and K+-dependent H+ fluxes in tonoplast vesicles isolated from storage tissue of red beet and sugar beet (Beta vulgaris L.). The Na+-dependent H+ flux across the tonoplast membrane could be resolved into two components: (a) a membrane potential-mediated flux through conductive pathways; and (b) an electroneutral flux which showed Michaelis-Menten kinetics relationship to Na+ concentration and was competitively inhibited by amiloride (Ki = 0.1 millimolar). The potential-dependent component of H+ flux showed an approximately linear dependence on Na+ concentration. In contrast, the K+-dependent H+ flux apparently consisted of a single component which showed an approximately linear dependence on K+ concentration, and was insensitive to amiloride. Based on the Na+- and K+-dependent H+ fluxes, the passive permeability of the vesicle preparation to Na+ was about half of that to K+.

The apparent Km for Na+ of the electroneutral Na+/H+ exchange varied by more than 3-fold (7.5-26.5 millimolar) when the internal and external pH values were changed in parallel. The results suggest a simple kinetic model for the operation of the Na+/H+ antiport which can account for the estimated in vivo accumulation ratio for Na+ into the vacuole.

1 Supported by the Natural Sciences and Engineering Research Council of Canada and the Department of Education of Quebec.




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