Abstract
Our previous investigations have established that Na+ translocation across the Tetraselmis viridis plasma membrane (PM) mediated by the primary ATP-driven Na+-pump, Na+-ATPase, is accompanied by H+ counter-transport [Y.V. Balnokin et al. (1999) FEBS Lett 462:402–406]. The hypothesis that the Na+-ATPase of T. viridis operates as an Na+/H+ exchanger is tested in the present work. The study of Na+ and H+ transport in PM vesicles isolated from T. viridis demonstrated that the membrane-permeant anion NO3 − caused (i) an increase in ATP-driven Na+ uptake by the vesicles, (ii) an increase in (Na++ATP)-dependent vesicle lumen alkalization resulting from H+ efflux out of the vesicles and (iii) dissipation of electrical potential, Δψ, generated across the vesicle membrane by the Na+-ATPase. The (Na++ATP)-dependent lumen alkalization was not significantly affected by valinomycin, addition of which in the presence of K+ abolished Δψ at the vesicle membrane. The fact that the Na+-ATPase-mediated alkalization of the vesicle lumen is sustained in the absence of the transmembrane Δψ is consistent with a primary role of the Na+-ATPase in driving H+ outside the vesicles. The findings allowed us to conclude that the Na+-ATPase of T. viridis directly performs an exchange of Na+ for H+. Since the Na+-ATPase generates electric potential across the vesicle membrane, the transport stoichiometry is mNa+/nH+, where m>n.
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Abbreviations
- BTP :
-
Bis-Tris-Propane, 1,3-bis[tris(hydroxymethyl)methylamino]-propane
- CCCP :
-
Carbonyl cyanide m-chlorophenylhydrazone
- DTT :
-
Dithiothreitol
- NCDC :
-
2-Nitro-4-carboxyphenyl N,N-diphenylcarbamate
- PMSF :
-
Phenylmethylsulfonyl fluoride
- PM :
-
Plasma membrane
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Acknowledgements
This work was supported by the Russian Foundation for Basic Research, Grant # 01-04-49135 and Grant # 02-04-06713. We thank Dr. A. Grabov (Imperial College, London) for critical reading of the manuscript and valuable comments.
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Balnokin, Y.V., Popova, L.G., Pagis, L.Y. et al. The Na+-translocating ATPase in the plasma membrane of the marine microalga Tetraselmis viridis catalyzes Na+/H+ exchange. Planta 219, 332–337 (2004). https://doi.org/10.1007/s00425-004-1224-7
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DOI: https://doi.org/10.1007/s00425-004-1224-7