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J. Biol. Chem., Vol. 265, Issue 17, 9617-9620, 06, 1990
S Blackford, PA Rea and D Sanders
The electrogenicity of H+/Ca2+ exchange in vacuolar membrane (tonoplast)
vesicles from Beta was studied to elucidate the role of this transport
system in vacuolar Ca2+ accumulation. To overcome the inherently high
proton permeability of tonoplast vesicles, the pH difference established by
the primary H(+)-ATPase was titrated to a uniform value by variation of the
concentration either of ATP or of a permanent anion (Cl-). This enabled
manipulation of membrane potential independently of the transmembrane pH
difference, with a higher inside- positive membrane potential produced at
lower Cl- concentrations. The rate and the extent of uncoupler-sensitive
Ca2+ uptake are both stimulated about 2-fold in conditions of more positive
membrane potential, suggesting that the transport system translocates
positive charge outward during Ca2+ uptake. A minimum integral H+:Ca2+
stoichiometry of 3 results in a driving force for Ca2+ accumulation in the
vacuole amounting to -140 mV in typical physiological conditions. It is
concluded that the antiporter is thermodynamically competent to account for
Ca2+ accumulation in plant vacuoles and that its reversal in vivo is
unlikely.
Voltage sensitivity of H+/Ca2+ antiport in higher plant tonoplast suggests a role in vacuolar calcium accumulation
Biology Department, University of York, Great Britain.
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