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Plant Physiology 82:121-125 (1986)
© 1986 American Society of Plant Biologists

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Articles

H+-Pumping Driven by the Plasma Membrane ATPase in Membrane Vesicles from Radish: Stimulation by Fusicoccin 1

Franca Rasi-Caldogno, Maria I. De Michelis, Maria C. Pugliarello and Erasmo Marrè

Centro di Studio del CNR per la Biologia Cellulare e Molecolare delle Piante, via G. Celoria 26, 20133 Milano, Italy, Dipartimento di Biologia "L. Gorini", via G. Celoria 26, 20133 Milano, Italy

The effect of fusicoccin on Mg:ATP-dependent H+-pumping in microsomal vesicles from 24-hour-old radish (Raphanus sativus L.) seedlings was investigated by measuring the initial rate of decrease in the absorbance of the {Delta}pH probe acridine orange. Fusicoccin stimulated Mg:ATP-dependent H+-pumping when the pH of the assay medium was in the range 7.0 to 7.6 while no effect of fusicoccin was detected between pH 6.6 and pH 6.0. Both basal and fusicoccin-stimulated H+-pumping were completely inhibited by vanadate and almost unaffected by nitrate. Fusicoccin did not change membrane permeability to protons and fusicoccin-induced stimulation of Mg:ATP-dependent H+-pumping was not affected by changes in the buffer capacity of the incubation medium. Deacetylfusicoccin stimulated H+-pumping as much as fusicoccin, while the physiologically inactive derivative 8-oxo-9-epideacetylfusicoccin did not. Stimulation of H+-pumping was saturated by 100 nanomolar fusicoccin. These data indicate that fusicoccin activates the plasma membrane H+-ATPase by acting at the membrane level independently of the involvement of other cell components. The percent stimulation by fusicoccin was the same at all ATP concentrations tested (0.5-5.0 millimolar), thus suggesting that with fusicoccin there is an increase in Vmax of the plasma membrane H+-ATPase rather than a decrease in its apparent Km for Mg:ATP.


1 Supported partially by the Research Project "Chimica Fine e Secondaria" (C-a) of the Italian Research Council.




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Copyright © 1986 by the American Society of Plant Biologists