Skip to main content
Log in

The proton gradient across the vacuo-lysosomal membrane of lutoids from the latex ofHevea brasiliensis. I. Further evidence for a proton-translocating ATPase on the vacuo-lysosomal membrane of intact lutoids

  • Articles
  • Published:
The Journal of Membrane Biology Aims and scope Submit manuscript

Summary

Lutoids (vacuo-lysosomal particles) were isolated from the latex ofHevea brasiliensis. Using flow dialysis with14C-methylamine uptake as a ΔpH probe and86Rb rubidium+valinomycin distribution for estimations of transmembrane electrical potential, intact lutoids exhibited a ΔpH of 1 unit (interior more acid) and a ΔΨ of −70 mV (interior negative), when suspended in an isotonic medium at physiological concentration of potassium (30mm) and pH 7.0, in the absence of ATP. In most cases, the Donnan potential was shown to fully account for ΔpH in nonenergized lutoids. The addition of Mg-ATP (5mm) resulted in a marked acidification of the lutoidic internal space (0.7 to 1 pH unit) depending on the composition of the medium, and in a membrane depolarization by 60 mV (interior becoming less negative). The resulting electrochemical potential of protons (\(\Delta \tilde \mu H\)) increased by a hundred millivolts when lutoids were energized by ATP. These data strongly support an inward electrogenic proton translocating function for the ATPase of the vacuo-lysosomal membrane of lutoids. Results are discussed in terms of thein vivo maintenance of large “lutoids/cytoplasm” proton gradients, and of the rôle of these vacuo-lysosomes in the homeostasis of the cytoplasmic metabolism.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Adams, H. 1965. Adénosine-5′ diphosphate and adenosine-5′-monophosphate.In: Methods of Enzymatic analysis. H.U. Bergmeyer, editor. pp. 573–580. Academic Press, New York

    Google Scholar 

  • Altendorf, K., Hiarata, H., Harol, F.M. 1975. Accumulation of lipid soluble ions and of rubidium as indicators of the electrical potential in membrane vesicles ofEscherichia coli.J. Biol. Chem 250:1405–1412

    PubMed  Google Scholar 

  • Archer, B.L., Barnard, D., Cockbain, E.G., Dickenson, P.B., McMullen, A.I. 1963. Structure, composition and biochemistry of latex.In: The Chemistry and Physics of Rubberlike Substances. pp. 41–72. McLaren & Sons, Ltd., London

    Google Scholar 

  • d'Auzac, J. 1975. Caractérisation d'une ATPase 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. 1977a. 1. Accumulation et pénétration du citrate et de la lysine dans les lutoïdes.In: Echanges ioniques Transmembranaires Chez les Végétaux.Colloq. C.N.R.S., Ed. CNRS Rouen, No. 258, pp. 392–398

  • d'Auzac, J., Dupont, J., Jacob, J.L., Lance, C., Marin, B., Moreau, F. 1977b. Un modèle de structure vacuolaire isolée intacte: Les lutoïdes du latex d'Hevea brasiliensis. 2. Caractéristique de la membrane lutoïdique.In: Echanges Ioniques Transmembranaires Chez les Végétaux.Colloq. C.N.R.S., Ed. CNRS Rouen, No. 258, pp. 399–406

  • d'Auzac, J., Lioret, C. 1974. Mise en évidence d'un mécanisme d'accumulation du citrate dans les lutoïdes du latex d'Hevea brasiliensis.Physiol. Veg. 12:617–635

    Google Scholar 

  • Brzozowska, J., Hanower, P., Chezeau, R. 1974. Free amino acids ofHevea brasiliensis latex.Experientia 30:894–895

    Google Scholar 

  • Brzozowska-Hanower, J., Cretin, H., Hanower, P., Michel, P. 1979. Variations de pH entre compartiments vacuolaire et cytoplasmique au sein du latex d'Hevea brasiliensis. Influence saisonnière et action du traitement par l'ethrel, générateur d'ethylène. Répercussion sur la production et l'apparition d'encoches sèches.Physiol. Veg. 17:889–905

    Google Scholar 

  • Collowick, S.P., Womack, F.C. 1969. Binding of diffusible molecules: Rapid measurement by rate of dialysis.J. Biol. Chem. 244:774–776

    PubMed  Google Scholar 

  • Coupe, M., Lambert, C. 1977. Absorption of citrate by the lutoids of latex, and rubber production by hevea.Phytochemistry 16:455–458

    Google Scholar 

  • Dell'Antone, P. 1979. Evidence for an ATP-driven “proton pump” in rate liver lysosomes by basic dye uptake.Biochem. Biophys. Res. Commun. 86:180–189

    PubMed  Google Scholar 

  • Dickenson, P.B. 1969. Electron microscopical studies of the latex vessels ofHevea brasiliensis.J. Rubber Res. Inst. Malays. 21:543–559

    Google Scholar 

  • Elema, R.P., Michels, P.A.M., Konings, W.N. 1978. Response of 9-amino-acridine fluorescence to transmembrane pH gradient in chromatophores from rhodopseudomonas spheroïdes.Eur. J. Biochem. 92:381–387

    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 

  • Henning, R. 1975. pH gradient across the lysosomal membrane generated by selective cation permeability and Donnan equilibrium.Biochim. Biophys. Acta 401:307–316

    PubMed  Google Scholar 

  • Hollemans, M., Donker-Koopman, W., Tager, J.M. 1980. A critical examination of the evidence for an Mg-ATP dependant proton pump in rat liver lysosomes.Biochim. Biophys. Acta 603:171–177

    PubMed  Google Scholar 

  • Jacob, J.L. 1970. Particularité de la glycolyse et de sa régulation au sein du latex d'Hevea brasiliensis.Physiol. Veg. 83:395–411

    Google Scholar 

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

    PubMed  Google Scholar 

  • Kell, B., John, P., Ferguson, S.J. 1978. The proton motive force in phosphorylating membrane vesicles fromParacocus denitrificans — magnitude, sites of generation and comparison with the phosphorylation potential.Biochem. J. 174:257–266

    PubMed  Google Scholar 

  • Lambert, C. 1975. Influence de l'ATP sur le pH intralutoïdique et la pénétration du citrate dans les lutoïdes du latex d'Hevea brasiliensis.C.R. Acad. Sci. Paris, t. 281-D:1705–1708

    Google Scholar 

  • Lin, W., Wanger, 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

    PubMed  Google Scholar 

  • Lowry, O.H., Rosebrough, N.J., Farr, A.L., Randall, R.J. 1951. Protein measurement with the Folin phenol reagent.J. Biol. Chem. 193:265–275

    PubMed  Google Scholar 

  • Marin, B. 1980. Some evidence about the occurrence of a Mg-ATP-dependant proton pump in plant vacuo-lysosomal compartment.In: Plant Membrane Transport: Current Conceptual Issues. R.M. Spanswick, W.J. Lucas, J. Dainty, editors. pp. 435–436. Elsevier-North Holland Biochemical, Amsterdam

    Google Scholar 

  • Marin, B. 1981. Le fonctionnement du transporteur tonoplastique de l'acide citrique du latex d'Hevea brasiliensis: Relation avec l'activité Adenosine. Triphosphatase membranaire. Ph.D. Thesis. Université des Sciences et techniques du Languedoc. Montpellier

  • Marin, B., Grignon. C. 1981. A computer-simulation of the mechanism of Citrate/H+ antiport at the tonoplast level in hevea latex.Can. J. Biochem. (Submitted)

  • Marin, B. Smith, J.A.C., Lüttge, M. 1981. The electrochemical proton gradient and its influence on citrate uptake in tonoplast vesicles ofHevea brasiliensis. Planta (in press)

  • McMullen, A.I. 1962. Particulate ribonucleoprotein components ofHevea brasiliensis latex.Biochem. J. 85:491–495

    PubMed  Google Scholar 

  • Mitchell, P., Moyle, J. 1968. Proton translocation coupled to ATP hydrolysis in rat liver mitochondria.Eur. J. Biochem. 4:530–539

    Google Scholar 

  • Moreau, F., Jacob, J.L., Dupont, J., Lance, C. 1975. Electron transport in the membrane of lutoids from the latex ofHevea brasiliensis.Biochim. Biophys. Acta 396:116–124

    PubMed  Google Scholar 

  • Padan, E., Rottenberg, H. 1973. Respiratory control and the proton electrochemical gradient in mitochondria.Eur. J. Biochem. 40:431–437

    Google Scholar 

  • Pujarniscle, S. 1968. Caractère lysosomal des lutoïdes du latex d'Hevea brasiliensis Müll Arg.Physiol. Veg. 6:27–46

    Google Scholar 

  • Pujarniscle, S. 1969. Etude de quelques facteurs intervenant sur la perméabilité et la stabilité de la membrane des lutoïdes d'Hevea brasiliensis.Physiol. Veg. 7:391–403

    Google Scholar 

  • Ramos, S., Schuldiner, S., Kaback, H.R. 1976. The electrochemical gradient of proton and its relationship to active transport inEscherichia coli membrane vesicles.Proc. Nat. Acad. Sci. USA 73:1892–1896

    PubMed  Google Scholar 

  • Reijngoud, D.J., Oud, P.S., Kas, J., Tager, J.M. 1976a. Relationship between medium pH and that of the lysosomal matrix as studied by two independant methods.Biomembranes 448:290–302

    Google Scholar 

  • Reijngoud, D.J., Oud, P.S., Tager, J.M. 1976b. Effects of ionophores on the intralysosomal pH.Biomembranes 448:303–313

    Google Scholar 

  • Reijngoud, D.J., Tager, L.M. 1973. Measurement of intralysosomal pH.Biochim. Biophys. Acta 297:174–178

    PubMed  Google Scholar 

  • Ribailler, D. 1972. Quelques aspects du rôle des lutoïdes dans la physiologie et l'écoulement du latex d'Hevea brasiliensis. Thèse Doc. Etat Sci. Nat. Abidjan, C.N.R.S.-AO-7716

  • Ribailler, D., Jacob, J.L., d'Auzac, J. 1971. Sur certains caractères vacuolaires des lutoïdes du latex d'Hevea brasiliensis.Physiol. Veg. 9:423–437

    Google Scholar 

  • Rottenberg, H., Grunwald, T., Avron, M. 1972. Determination of pH in chloroplasts: I. Distribution of14C-methylamine.Eur. J. Biochem. 25:54–63

    PubMed  Google Scholar 

  • Ruinen, J. 1950. Microscopy of the lutoids in hevea latex.Ann. Bogor. I:27–45

    Google Scholar 

  • Schneider, D.L. 1977. Membranous localization and properties of ATPase of rat liver lysosomes.J. Membrane Biol. 34:247–261

    Google Scholar 

  • Schneider, D.L. 1979. The acidification of rat liver lysosomesin vitro. A role for the membranous ATPase as a proton pump.Biochem. Biol. Res. Commun. 86:180–189

    Google Scholar 

  • Sorgato, M., Ferguson, S.J., Kell, D.B., John, P. 1978. The proton motive force in bovine heart submitochondrial particles; magnitude, site of generation and comparison with the phosphorylation potential.Biochem. J. 174:237–256

    PubMed  Google Scholar 

  • Taysum, D.H. 1960. The establishment of a bacterial population in latex vessels during normal tapping.Proc. Nat. Rubber Res. Conf., Kuala-Lumpur., pp. 856–871

  • Tupy, J., 1973. The activity of latex invertase and latex production inHevea brasiliensis Müll. Arg.Physiol. Veg. 11:633–641

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Cretin, H. The proton gradient across the vacuo-lysosomal membrane of lutoids from the latex ofHevea brasiliensis. I. Further evidence for a proton-translocating ATPase on the vacuo-lysosomal membrane of intact lutoids. J. Membrain Biol. 65, 175–184 (1982). https://doi.org/10.1007/BF01869961

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01869961

Key words

Navigation