Changes in the ordering of lipids in the membrane of Dunaliella in response to osmotic-pressure changes. An e.s.r. study.

Changes in the ordering and motion of lipids in response to changes in the external solute concentration have been studied by using the 5-nitroxide stearate (5NS) and 16-nitroxide stearate (16NS) spin probes in the plasma membrane of the halotolerant unicellular alga Dunaliella salina. Increases in ordering of the 5NS probe and decreases in motion of the 16NS probe were observed in cells equilibrated over 18 h at increasing NaCl concentrations. These changes probably resulted from the influence of the high NaCl concentration on the charged phospholipid head groups of the membrane. A short-term (less than 100 min) decrease in the order parameter, S, of the 5NS probe was observed for cells swollen by exposure to a sudden decrease of NaCl concentration from 5.0 to 2.5 M. After 100 min the value of S for 5NS was close to the value obtained in cells that had been equilibrated in 2.5 M-NaCl for 18 h. Since the cells had regained their original size and shape by 100 min it was assumed that the short-term decrease in S was associated with the swelling. A similar result was obtained when the cells were suddenly changed from 3.0 M- to 1.5 M-sorbitol. Conversely, an increase in S was observed for cells shrunk when the external solute concentration was doubled from 1.5 M- to 3.0 M-NaCl. As the cells regained their original size and shape the value of S decreased to the value observed in cells that had been equilibrated in 3.0 M-NaCl for 18 h. It is suggested that the changes in S are related to the movement of lipid into or out of a reservoir of membrane material as the membrane shrinks or expands. This movement of lipid maintains the tension of the membrane below the value at which it is disrupted. Such changes in lipid ordering could provide a mechanism whereby information about external osmotic-pressure changes is transmitted across the cell wall.