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Electrostatic surface properties of plasmalemma vesicles from oat and wheat roots. Ion binding and screening investigated by 9-aminoacridine fluorescence

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Abstract

Right-side-out and sealed plasmalemma vesicles were isolated from roots of spring wheat (Triticum aestivum L. cv. Drabant) and oat (Avena sativa L. cv. Brighton) by two-phase partition in a medium containing sucrose (0.25 mol l-1). Oat root plasmalemma vesicles were discovered to contain a strongly fluorescent compound with an emission maximum at 418 nm. The surface potential of the membranes was monitored by 9-aminoacridine fluorescence and the effect of protein concentration, mannitol versus sucrose, absence of osmoticum, concentrations of salt, and titrations with chelators investigated. It is concluded that i) protein concentrations of less than 50 μg ml-1 for oat and 100 μg ml-1 for wheat plasmalemma vesicles should be used to avoid serious problems with non-linearity of response of 9-aminoacridine fluorescence, ii) mannitol can be used instead of sucrose as the osmoticum, iii) the vesicles were ruptured in the absence of osmoticum allowing us to monitor both sides of the membranes, iv) plasmalemma vesicles from oat roots are more negative than vesicles from wheat roots, and v) oat and wheat root plasmalemma vesicles are isolated with about the same amounts of bound Ca2+ and Mg2+. These bound divalent cations may not, however, reflect the in-vivo conditions since the tissues were homogenised in the presence of ethylenediaminetetraacetic acid.

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Abbreviations

EDTA:

ethylenediaminetetraacetic acid

EGTA:

ethylene glycol-bis(β-aminoethyl ether)-N,N,N′,N′-tetraacetic acid

c1/2 value:

concentration at which half of the maximum effect is observed

Mops:

3-(N-morpholino)propanesulfonic acid

References

  1. Åkerlund, H.-E., Andersson, B., Persson, A., Albertsson, P.-0A. (1979) Isoelectric points of spinach thylakoid membrane surfaces as determined by cros partition. Biochim. Biophys. Acta 552, 238–246

  2. Barber, J. (1980) Membrane surface charges and potentials in relation to photosynthesis. Biochim. Biophys. Acta 594, 253–308

  3. Barber, J. (1982) Influence of surface charges on thylakoid structure and function. Annu. Rev. Plant Physiol. 33, 261–295

  4. Bérczi, A., Møller, I.M., Oláh, Z., Lundborg, T., Erdei, L. (1984a) A model for cation content of plants based on surface potentials and surface charge densities of plant membranes. Physiol. Plant. 61, 529–534

  5. Bérczi, A., Møller, I.M., Lundborg, T., Kylin, A. (1984b) The surface charge density of wheat root membranes. Physiol. Plant. 61, 535–540

  6. Douzou, P., Maurel, P. (1977) Ionic control of biochemical reactions. Trends Biochem. Sci. 2, 14–17

  7. Ericson, I. (1974) Determination of the isoelectric point of rat liver mitochondria by cross-partition. Biochim. Biophys. Acta 356, 100–107

  8. Gibrat, R., Grignon, C. (1982) Effect of pH on the surface charge density of plant membranes. Comparison of microsomes and liposomes. Biochim. Biophys. Acta 692, 462–468

  9. Glass, A.D.M. (1976) Regulation of potassium absorption in barley roots. An allosteric model. Plant Physiol. 58, 33–37

  10. Hackett, D.P. (1961) Effects of salts on DPNH oxidase activity and structure of sweet potato mitochondria. Plant Physiol. 36, 445–452

  11. Jensén, P., Pettersson, S. (1978) Allosteric regulation of potassium uptake in plant roots. Physiol. Plant. 42, 207–213

  12. Johnston, S.P., Møller, I.M., Palmer, J.M. (1979) The stimulation of exogenous NADH oxidation in Jerusalem artichoke mitochondria by screening of charges on the membranes. FEBS Lett. 108, 28–32

  13. Kähr, M., Møller, I.M. (1976) Temperature response and effect of Ca2+ and Mg2+ on ATPases from roots of oats and wheat as influenced by growth temperature and nutritional status. Physiol. Plant. 38, 153–158

  14. Körner, L., Møller, I.M., Kjellbom, P., Larsson, C. (1984) Surface properties of plasmalemma vesicles from barley roots and shoots. In: 4th Congress of FESPP, Strasbourg, France, 29 July–3 August 1984, pp. 515–516

  15. Larsson, C., Kjellbom, P., Widell, S., Lundborg, T. (1984) Sidedness of plant plasma membrane vesicles purified by partition in aqueous two-phase systems. FEBS Lett. 171, 271–276

  16. 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

  17. Lundborg, T., Widell, S., Bérczi, A., Kjellbom, P., Larsson, C., Møller, I.M. (1984) Sidedness and surface charge density of plasmalemma vesicles. In: 4th Congress of FESPP, Strasbourg, France, 29 July–3 August 1984, pp. 523–524

  18. Lundborg, T., Widell, S., Larsson, C. (1981) Distribution of ATPases in wheat root membranes separated by phase partition. Physiol. Plant 52, 89–95

  19. Maurel, P., Douzou, P. (1976) Catalytic implications of electrostatic potentials.: The lytic activity of lysozyme as a model. J. Mol. Biol. 102, 253–264

  20. McLaughlin, S. (1977) Electrostatic potentials at membrane-solution interfaces. Curr. Top. Membr. Transp. 9, 71–143

  21. Møller, I.M. (1983) Monitoring of membrane-bound divalent cations in plant mitochondria using chlorotetracycline fluorescence. Physiol. Plant. 59, 567–572

  22. Møller, I.M., Chow, W.-S., Palmer, J.M., Barber, J. (1981a) 9-Aminoacridine as a fluorescent probe of the electrical diffuse layer associated with the membranes of plant mitochondria. Biochem. J. 193, 37–46

  23. Møller, I.M., Johnston, S.P., Palmer, J.M. (1981b) A specific role for Ca2+ in the oxidation of exogenous NADH by Jerusalem artichoke (Helianthus tuberosus) mitochondria. Biochem. J. 194, 487–495

  24. Møller, I.M., Kay, C.J., Palmer, J.M. (1984a) Electrostatic screening stimulates rate-limiting steps in mitochondrial electron transport. Biochem. J. 223, 761–767

  25. Møller, I.M., Lundborg, T. (1985) A fluorescent compound in oat root plasma membrane. Physiol. Plant., in press

  26. Møller, I.M., Lundborg, T., Bérczi, A. (1984b) The negative surface charge density of plasmalemma vesicles from wheat and oat roots. FEBS Lett. 167, 181–185

  27. Møller, I.M., Palmer, J.M. (1981) Charge screening by cations affects the conformation of the mitochondrial inner membrane. A study of exogenous NAD(P)H oxidation in plant mitochondria. Biochem. J. 195, 583–588

  28. Møller, I.M., Schwitzguébel, J.-P., Palmer, J.M. (1982) Binding and screening by cations and the effect on exogenous NAD(P)H oxidation in Neurospora crassa mitochondria. Eur. J. Biochem. 123, 81–88

  29. Moore, A.L., Åkerman, K.E.O. (1982) Ca2+ stimulation of the external NADH dehydrogenase in Jerusalem artichoke (Helianthus tuberosus) mitochondria. Biochem. Biophys. Res. Commun. 109, 513–517

  30. Nicholls, P. (1974) Cytochrome c binding to enzymes and membranes. Biochim. Biophys. Acta 346, 261–310

  31. Reed, K.C., Bygrave, F.L. (1974) Accumulation of lanthanum by rat liver mitochondria. Biochem. J. 138, 239–252

  32. Sack, F.D., Priestley, D.A., Leopold, A.C. (1983) Surface charge on isolated maize-coleoptile amyloplasts. Planta 157, 511–517

  33. Schuldiner, S., Rottenberg, H., Avron, M. (1972) Determination of ΔpH in chloroplasts. 2. Fluorescent amines as a probe for the determination of ΔpH in chloroplasts. Eur. J. Biochem. 25, 64–70

  34. Searle, G.F.W., Barber, J., Mills, J.D. (1977) 9-Amino-acridine as a probe of the electrical double layer associated with the chloroplast thylakoid membranes. Biochim. Biophys. Acta 461, 413–425

  35. Sommarin, M., Lundborg, T., Kylin, A. (1984) Oat and wheat root plasmalemma purified by two-phase partitioning —ATPase activities, specificities and inhibitions. In: 4th Congress of FESPP, Strasbourg, France, 29 July–3 August 1984, pp. 571–572

  36. Telfer, A., Barber, J., Jagendorf, A.T. (1980) Electrostatic control of chloroplast coupling factor binding to thylakoid membranes as indicated by cation effects on electron transport and reconstitution of photophosphorylation. Biochim. Biophys. Acta 591, 331–345

  37. Thibaud, J.-B., Romieu, C., Gibrat, R., Grouzis, J.-P., Grignon, C. (1984) Local ionic environment of plant membranes: Effects of membrane functions. Z. Pflanzenphysiol. 114, 207–213

  38. Wehrle, J.P., Jurkowitz, M., Scott, K.M., Brierley, G.P. (1976) Mg2+ and the permeability of heart mitochondria to monovalent cations. Arch. Biochem. Biophys. 174, 312–323

  39. Weinstein, J.N., Blumenthal, R., van Renswoude, J., Kempf, C., Klausner, R.D. (1982) Charge clusters and the orientation of membrane proteins. J. Membr. Biol. 66, 203–212

  40. Widell, S., Lundborg, T., Larsson, C. (1982) Plasma membranes from oats prepared by partition in an aqueous polymer two-phase system. On the use of light-induced cytochrome b reduction as a marker for the plasma membrane. Plant Physiol. 70, 1429–1435

  41. Wignarajah, K., Lundborg, T., Björkman, T., Kylin, A. (1983) Field properties and ion uptake of wheat and oat: are they expressed in the modulatory influences of ions on membrane ATPases? Oikos 40, 6–13

  42. Wojtczak, L., Nalecz, M.J. (1979) Surface charge of biological membranes as a possible regulator of membrane-bound enzymes. Eur. J. Biochem. 94, 99–107

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Møller, I.M., Lundborg, T. Electrostatic surface properties of plasmalemma vesicles from oat and wheat roots. Ion binding and screening investigated by 9-aminoacridine fluorescence. Planta 164, 354–361 (1985). https://doi.org/10.1007/BF00402946

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Key words

  • 9-Aminoacridine
  • Avena (plasmalemma)
  • Bound divalent cations
  • Plasmalemma (electrostatic surface properties)
  • Triticum (plasmalemma)