Effects of ethanol on rat brain (Na + K)ATPase from native and delipidized synaptic membranes

Abstract

The role of lipids in the effect of ethanol on synaptosomal (Na + K)ATPase was studied using native and partially delipidized synaptosomal membranes from control and alcoholic rats. A biphasic effect of alcohol was observed with the (N + K)ATPase from control membranes. Ethanol at low concentrations (< 100 mM) appears to enhance the enzyme activity, but at higher concentrations (> 300 mM) was inhibitory. The biphasic response to ethanol was also observed with the (Na + K)ATPase isolated from alcoholic animals; however, in this case the enzyme showed a resistance to the inhibitory effect of ethanol. Delipidization of synaptic membranes with Lubrol WX or phospholipase A practically abolishes the effects of alcohol on (Na + K)ATPase from both control and alcoholic animals. It thus seems that the effects of ethanol are due mainly to their interaction with the lipids surrounding the enzyme. Furthermore, addition of ethanol to native membranes did not change the V max and K m for K +. However, when ethanol at the same concentration was added to delipidized membranes, a decrease in K m with no change in V max was observed. Ethanol under these conditions apparently interacts also with the enzyme protein. On the other hand, chronic ethanol intake produces an increase of both V max and K m for K +. However, when alcohol was added in vitro, there were no changes in the kinetic parameters of either native or delipidized membranes. These data indicate that although the effects of ethanol on synaptosomal (Na + K)ATPase are mainly due to its interaction with the lipid microenvironment of the enzyme, a direct ethanol action on the enzyme protein also occurs. Our data further suggest that chronic ethanol treatment alters enzyme sensitivity to the effect of ethanol which may be related to the membrane-lipid composition and/or to changes in the conformation of the enzyme protein.