Brain (Na+,K+)-ATPase. Opposite effects of ethanol and dimethyl sulfoxide on temperature dependence of enzyme conformation and univalent cation binding.
We examined effects of ethanol and dimethyl sulfoxide on the regulation and apparent thermodynamic properties of moderate affinity Na+ and K+ binding that regulates the K+-dependent phosphatase activity of (Na+,K+)-ATPase. Ethanol and other alcohols reduced the apparent affinity for Na+ and K+ at their moderate affinity sites and increased the negative delta H and delta S of cation binding. Dimethyl sulfoxide had the opposite effects. Inhibition by ethanol was favored by high temperature or low K+. Ethanol potentiated inhibition of K+ binding by ATP or Mg2+. Ethanol also shifted the equilibrium between K+-sensitive and -insensitive forms of (Na+,K+)-ATPase toward the K+-sensitive form; in this case, it reduced the negative delta H and delta S for the transition to K+-sensitive enzyme. Again, dimethyl sulfoxide had the opposite effects. These data indicate that ethanol and other agents considered to affect membrane fluidity act by a combination of membrane (on cation binding) and solvent (on conformation) effects. The most important effect of ethanol and similar agents on the enzyme is to prevent the formation of K+-sensitive enzyme by cation binding and to destabilize K+-sensitive enzyme in the presence of ATP. These results also add further evidence that the sites by which Na+ and K+ produce K+-sensitive enzyme are similar or identical.
D000438:Alcohols / Q000494:pharmacology; D000818:Animals; D001921:Brain / Q000201:enzymology*; D002414:Cations, Monovalent; D002417:Cattle; D004121:Dimethyl Sulfoxide / Q000494:pharmacology*; D000431:Ethanol / Q000494:pharmacology*; D007700:Kinetics; D008024:Ligands; D011487:Protein Conformation / Q000187:drug effects; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism*; D013696:Temperature
D000438:Alcohols; D002414:Cations, Monovalent; D008024:Ligands; D000431:Ethanol; D000254:Sodium-Potassium-Exchanging ATPase; D004121:Dimethyl Sulfoxide