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Evidence for an electrogenic adenosine-triphosphatase in Hevea tonoplast vesicles

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Abstract

The function of the Mg-dependent ATPase of Hevea tonoplast in active proton transport was investigated by using a purified tonoplast fraction containing tightly sealed vesicles. In the used experimental conditions, the uptake of [14C]triphenylmethyl-phosphonium ion ([14C]TPMP+) and [3H]tetraphenyl-phosphonium ion ([3H]TPP+) by the vesicles indicated a transmembrane potential difference, negative inside. In parallel, the uptake of [14C]methylamine into the vesicles monitored a transmembrane pH gradient, interior acid. The addition of 5 mM Mg-ATP markedly depolarized the membrane and increased the magnitude of trnasmembrane pH gradient. These ATP-driven events were substrate specific for Mg-ATP. They were strongly inhibited by ATPase inhibitors such as N, N′-dicyclohexylcar-bodiimide. They were completely eliminated by proton conductors such as carbonylcyanide-p-trifluoromethoxy-phenylhydrazone and 5-chloro-3-tert-butyl-2′-chloro-4-nitro-salicylanilide. They depended on the pH of the medium, the maximum being reached at about pH 7.0. These data provide in vitro evidence that the Mg-ATPase localized at tonoplast level is an electrogenic pump. They are consistent with the hypothesis that an electrogenic H+ pump is catalyzed by the tonoplast ATPase of higher plants.

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Abbreviations

ΔpH:

transmembrane pH gradient

Δψ:

electrical potential difference across membrane

DCCD:

N, N′-dicyclohexyl-carbodiimide

FCCP:

carbonyl-cyanide-p-trifluoromethoxy-phenylhydrazone

clone PB 86:

clone Prang Besar 86

S-13:

5-chloro-3-tert-butyl-2′-chloro-4-nitro-salicylanilide

TPMP+ :

triphenyl-methyl-phosphonium ion

TPP+ :

tetraphenylphosphonium ion

References

  • d'Auzac, J. (1975) Caractérisation d'une ATP-ase membranaire en présence d'une phosphatase acide dans les lutoïdes du latex d'Hevea brasiliensis. Phytochemistry 14, 671–675

    Google Scholar 

  • d'Auzac, J. (1977) ATPase membranaire de vacuoles lysosomales: les lutoïdes du latex d'Hevea brasiliensis. Phytochemistry 16, 1881–1885

    Google Scholar 

  • d'Auzac, J., Brzozowska, J., Hanower, P., Lambert, C., Lioret, C., Niamien N'Goran, M. (1977) Un modèle de structure vacuolaire isolée intacte: les lutoïdes du latex d'Hevea brasiliensis. I. Accumulation et pénétration du citrate et de la L-lysine dans les lutoïdes. In: Transmembrane ionic exchanges in plants, pp. 391–398, Thellier, M., Monnier, A., Demarty, M., Dainty, J. eds. Editions du Centre National de la Recherche Scientifique.; Publications de l'Université de Rouen, Rouen

    Google Scholar 

  • d'Auzac, J., Crétin, H., Marin, B., Lioret, C. (1982) A plant vacuolar system: the lutoïds from Hevea brasiliensis latex. Physiol. Vég. 20, 311–331

    Google Scholar 

  • Crétin, H. (1982) The proton gradient across the vacuo-lysosomal membrane of lutoïds from the latex of Hevea brasiliensis. I. Further evidence for a proton-translocating ATPase on the vacuo-lysosomal membrane of intact lutoïds. J. Membr. Biol. 65, 175–184

    Google Scholar 

  • Crétin, H., Marin, B., d'Auzac, J. (1982) Characterization of a magnesium-dependent proton translocating ATPase on Hevea latex tonoplast. In: Plasmalemma and tonoplast: their functions in the plant cell, pp. 201–208, Marmé D., Marré, E., Hertel, R., eds.). Elsevier Biomedical Press, Amsterdam New York Oxford

    Google Scholar 

  • Goffeau, A., Slayman, C.W. (1981) The proton-translocating ATPase of the fungal plasma membrane. Biochim. Biophys. Acta 639, 197–223

    Google Scholar 

  • Hager, A., Frenzel, R., Laible, D. (1980) ATP-dependent proton transport into vesicles of microsomal membranes of Zea mays coleoptiles. Z. Naturforsch. 35c, 783–793

    Google Scholar 

  • Hager, A., Helmle, M. (1981) Properties of an ATP-fueled, Cl− dependent proton pump localized in membranes of microsomal vesicles from maize coleoptiles. Z. Naturforsch. 36c, 997–1008

    Google Scholar 

  • Hanower, P., Brzozowska, J., Niamien N'Goran, M. (1977) Absorption des acides aminés par les lutoïdes du latex d'Hevea brasiliensis. Physiol. Plant. 39, 299–304

    Google Scholar 

  • Jacob, J.L., Sontag, N. (1974) Purification et étude de la phosphatase acide lutoïdique du latex d'Hevea brasiliensis. Biochimie 56, 1315–1322

    Google Scholar 

  • Johnson, R.G., Scarpa, A. (1976) Internal pH of isolated chromaffin vesicles. J. Biol. Chem. 251, 2189–2191

    Google Scholar 

  • Kakinuma, Y., Ohsumi, Y., Anraku, Y. (1981) Properties of H+-translocating adenosine-triphosphatase in vacuolar membranes of Saccharomyces cerevisiae. J. Biol. Chem. 256, 10859–10863

    Google Scholar 

  • Komor, E., Tanner, W. (1976) The determination of the membrane potential of Chlorella vulgaris. Evidence for electrogenic sugar transport. Eur. J. Biochem. 70, 197–204

    Google Scholar 

  • Lambert, C. (1975) Influence de l'ATP sur le pH intralutoïdique et sur la pénétration du citrate dans les lutoïdes du latex d'Hevea brasiliensis. C.R. Acad. Sci. Ser. D 281, 1705–1708

    Google Scholar 

  • Lin, W., Wagner, G.J., Siegelman, H.W., Hind, G. (1977) Membrane-bound ATPase of intact vacuoles and tonoplasts isolated from mature plant tissue. Biochim. Biophys. Acta 465, 110–117

    Google Scholar 

  • Lowry, O.H., Rosebrough, N.J., Farr, A.L., Randall, R.J. (1951) Protein measurements with the Folin phenol reagent. J. Biol. Chem. 193, 265–1275

    Google Scholar 

  • Makkar, H.P.S., Sharma, O.P., Negi, S.S. (1980) Assay of proteins by Lowry's method in the presence of high concentrations of β-mercaptoethanol. Anal. Biochem. 104, 124–126

    Google Scholar 

  • Marin, B. (1982) Le fonctionnement du transporteur tonoplastique du citrate du latex d'Hevea brasiliensis. Trav. Doc. O.R.S.T.O.M. 144, 1–409

    Google Scholar 

  • Marin, B., Blasco, F. (1982) Further evidence for the proton pumping work of tonoplast ATPase from Hevea latex. Biochem. Biophys. Res. Commun. 105, 354–361

    Google Scholar 

  • Marin, B., Cretin, H., d'Auzac, J. (1982a) Energization of solute transport and accumulation at tonoplast level in Hevea latex. Physiol. Vég. 20, 333–346

    Google Scholar 

  • Marin, B., Cretin, H., d'Auzac, J. (1982b) Energization of solute transport and accumulation in the Hevea latex vacuole. In: Plasmalemma and tonoplast: their functions in the plant cell, pp. 209–216, Marmé, D., Marré, E., Hertel, R., eds. Elsevier Biomedical Press, Amsterdam New York Oxford

    Google Scholar 

  • Marin, B., Marin-Lanza, M., Komor, E. (1981 a) The protonmotive potential difference across the vacuo-lysosomal membrane of Hevea brasiliensis and its modification of a membrane-bound adenosine triphosphatase. Biochem. J. 198, 365–372

    Google Scholar 

  • Marin, B., Smith, J.A.C., Lüttge, U. (1981 b) The protonmotive force and its influence on citrate uptake in vacuolar vesicles of Hevea brasiliensis latex. Planta 153, 486–493

    Google Scholar 

  • Marin, B., Trouslot, P. (1975) The occurrence of ribonucleic acid in the lutoïd fraction (lysosomal compartment) from Hevea brasiliensis Künth. (Müll.-Arg.). latex. Planta 124, 31–41

    Google Scholar 

  • McRobbie, E.A.C. (1975) Ion transport in plant cells. Curr. Top. Membr. Transp. 7, 1–48

    Google Scholar 

  • Mettler, I.J., Leonard, R.T. (1979) Isolation and partial characterization of vacuoles from tobacco protoplasts. Plant Physiol. 64, 1114–1120

    Google Scholar 

  • Poole, R.J. (1978) Energy coupling for membrane transport. Annu. Rev. Plant Physiol. 29, 437–460

    Google Scholar 

  • Rasi-Caldogno, F., De Michelis, M.I., Pugliarello, M.C. (1981) Evidence for an electrogenic ATPase in microsomal vesicles from pea internodes. Biochim. Biophys. Acta 642, 27–45

    Google Scholar 

  • Raven, J.A., Smith, F.A. (1979) Intracellular pH and its regulation. Annu. Rev. Plant Physiol. 30, 289–311

    Google Scholar 

  • Rottenberg, H. (1979) The measurement of membrane potential and ΔpH in cells, organelles and vesicles. Methods Enzymol. 55, 547–569

    Google Scholar 

  • Smith, F.A., Raven, J.A. (1976) Chloride transport in Characorallina and the electrochemical potential difference for hydrogen ions. J. Exp. Bot. 27, 451–459

    Google Scholar 

  • Stout, R.G., Cleland, R.E. (1982) Evidence for a Cl−-stimulated Mg ATPase proton pump in oat root membranes. Plant Physiol. 69, 798–803

    Google Scholar 

  • Sze, H., Churchill, K.A. (1981) Mg2+/KCl-ATPase of plant membranes is an electrogenic pump. Proc. Natl. Acad. Sci. USA 78, 5578–5582

    Google Scholar 

  • Taussky, H.H., Shorr, E. (1953) A microcolorimetric method for the determination of inorganic phosporus. J. Biol. Chem. 202, 675–685

    Google Scholar 

  • Wagner, G.J. (1981) Enzymic and protein character of tonoplast from Hippeastrum vacuoles. Plant Physiol. 68, 499–503

    Google Scholar 

  • Walker, R.R., Leigh, R.A. (1981) Characterisation of a saltstimulated ATPase activity associated with vacuoles isolated from storage roots of red beet (Beta vulgaris L.). Planta 153, 140–149

    Google Scholar 

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  1. Bernard Marin

    Present address: Botanisches Institut der Universität, Universitätsstrasse 30, D-8580, Bayreuth, Germany

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  1. Laboratoire de Physiologie Végétale Appliquée, Université des Sciences et Techniques du Languedoc, Place Eugéne Bataillon, F-34060, Montpellier-Cédex, France

    Bernard Marin

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Marin, B. Evidence for an electrogenic adenosine-triphosphatase in Hevea tonoplast vesicles. Planta 157, 324–330 (1983). https://doi.org/10.1007/BF00397403

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  • DOI: https://doi.org/10.1007/BF00397403

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