Abstract
A highly enriched plasma membrane fraction has been isolated from dark-grown cotyledons ofRicinus communis by phase partitioning. This is demonstrated by the properties of the associated ATPase: high vanadate sensitivity, azide and nitrate insensitivity, sharp pH optimum around 6.5, and high specificity for ATP as substrate. The upper plasma membrane fraction also contained a pyrophosphatase activity, normally considered to be located on the tonoplast or Golgi membranes, which showed a specific activity higher than that in the lower phase. Sucrose gradient centrifugation of both microsomal and upper phase fractions showed a comigration of some pyrophosphatase activity with the plasma membrane fraction. Sucrose uptake changes with development inRicinus cotyledons. The ATPase activity in the upper (plasma membrane) phase also varied in a similar way with development, whereas activity in the lower phase showed little change. Pyrophosphatase activity in the upper phase also increased with development but did not show a peak and fall as seen for sucrose uptake and ATPase. The possibility that changes in plasma membrane ATPase may contribute to changes in sucrose uptake capacity and the possible cellular origin and physiological significance of the pyrophosphatase activity are discussed.
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Abbreviations
- Pi:
-
orthophosphate
- PPase:
-
pyrophosphatase
- PPi:
-
pyrophosphate; SW26, 2,2,2-trichloroethyl 3,4-dichloro-carbanilate
References
Bradford M.M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem.72, 248–254
Chanson A., McNaughton E., Taiz L. (1984) Evidence for a KCl-stimulated, Mg2+ ATPase on the Golgi of corn coleoptiles. Plant Physiol.76, 498–507
Chanson A., Fichmann J., Spear D., Taiz L. (1985) Pyrophosphate-driven proton transport by microsomal membranes of corn coleoptiles. Plant Physiol.79, 159–164
Giaquinta R.T. (1983) Phloem loading of sucrose. Annu. Rev. Plant Physiol.34, 347–387
Gross P., ap Rees T. (1986) Akaline inorganic pyrophosphatase and starch synthesis in amyloplasts. Planta167, 140–145
Hodges, T.K., Leonard, R.T. (1972) Purification of a plasma membrane-bound adenosine triphosphatase from plant roots. Methods Enzymol32, 392–406
Humphreys, T.E. (1988) Phloem transport with emphasis on loading and unloading. In: Solute transport in plant cells and tissues pp. 305–345, Baker D.A., Hall, J.L., eds. Longman, Harlow
Hutchings V.M. (1978) Sucrose and proton cotransport inRicinus cotyledons. Planta138, 229–235
Komor E. (1977) Sucrose uptake by cotyledons ofRicinus communis L.: characteristics, mechanism and regulation. Planta137, 119–131
Komor E., Rotter M., Tanner W. (1977) A proton-cotransport system in a higher plant: sucrose transport inRicinus communis. Plant Sci. Lett.9, 153–162
Larsson C. (1983) Partition in aqueous polymer two-phase systems. In: Isolation of membranes and organelles from plant cells, pp. 277–309, Hall, J.L., Moore, A.C., eds. Academic Press, London
McCarty D.R., Keegstra K., Selman B.R. (1984) Characterization and localisation of the ATPase associated with pea chloroplast envelope membranes. Plant Physiol.76, 584–588
Ohnishi T., Gall R.S., Mayer M.L. (1975) An improved assay of inorganic phosphate in the presence of extralabile phosphate compounds: application to the ATPase assay in the presence of phosphocreatine. Anal. Biochem.69, 261–267
Petel G., Gendraud M. (1989) Localization in sucrose gradients of pyrophosphatase activities in the microsomal fraction of Jerusalem artichoke (Helianthus tuberosus L.) tubers. J. Plant Physiol.134, 466–470
Poole R.J. (1988) Plasma membrane and tonoplast. In: Solute transport in plant cells and tissues, pp 83–105, Baker D.A., Hall J.L., eds. Longman, Harlow
Vreugdenhil D., Spanswick R.M. (1987) The effect of vanadate on proton-sucrose cotransport inRicinus cotyledons. Plant Physiol.84, 605–608
Wightman F., Forest J.C. (1978) Properties of plant aminotransferases. Phytochemistry17, 1455–1471
Williams L., Hall J.L. (1987) ATPase and proton pumping activities in cotyledons and other phloem-containing tissues ofRicinus communis. J. Exp. Bot.38, 185–202
Williams L.E., Hall J.L. (1989) ATPase and proton-translocating activities in a plasma membrane-enriched fraction from cotyledons ofRicinus communis. J. Exp. Bot.40, 1205–1213
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We would like to thank the Science and Engineering Research Council (UK) for financial support.
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Williams, L.E., Nelson, S.J. & Hall, J.L. Characterization of solute transport in plasma membrane vesicles isolated from cotyledons ofRicinus communis L.. Planta 182, 532–539 (1990). https://doi.org/10.1007/BF02341028
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DOI: https://doi.org/10.1007/BF02341028