CHARACTERIZATION OF SOLUTE TRANSPORT IN PLASMA-MEMBRANE VESICLES ISOLATED FROM COTYLEDONS OF RICINUS-COMMUNIS L .1. ADENOSINE-TRIPHOSPHATASE AND PYROPHOSPHATASE ACTIVITIES ASSOCIATED WITH A PLASMA-MEMBRANE FRACTION ISOLATED BY PHASE PARTITIONING

A highly enriched plasma membrane fraction has been isolated from dark-grown cotyledons of Ricinus 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 in Ricinus 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. © 1990 Springer-Verlag.