Lipid requirement of the membrane sodium-plus-potassium ion-dependent adenosine triphosphatase system.
The dependence of the (Na-++K-+)-dependent ATPase (adenosine triphosphatase) (EC 3.6.1.3) on lipid has been examined in a number of different ways, with the use of various preparations from kidney tissue. The main findings were as follows. (1) The ATPase activities of the preparations examined were closely correlated with their total phospholipid content. (2) Extraction of the ATPase with deoxycholate or Lubrol W, combined with suitable salt-fractionation and washing procedures, removed phospholipid, cholesterol and enzymic activity in parallel; but activity was completely lost before all lipid had been removed. (3) The loss of activity could not be attributed to inhibition by residual detergent. (4) No selective removal of any particular phospholipid class by detergent could be detected. (5) Consistent reactivation of the Lubrol-extracted enzymes was obtained  by adding dispersions of exogenous phospholipid, but only some, bearing a net negative charge, such as phosphatidylserine and phosphatidylglycerol, were effective. (6) The degree of reactivation was correlated with the amount of residual activity remaining after lipid depletion. (7) Partial purification of the ATPase, giving a 50-fold increase in specific activity, was not accompanied by selective enhancement of any particular class of phospholipid. We conclude that although the ATPase is dependent on phospholipid, only the reactivation results provide evidence for specificity.
D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000818:Animals; D002458:Cell Fractionation; D002850:Chromatography, Gel; D003840:Deoxycholic Acid; D003902:Detergents; D007668:Kidney / Q000201:enzymology / Q000648:ultrastructure*; D007700:Kinetics; D050356:Lipid Metabolism; D008566:Membranes / Q000201:enzymology / Q000378:metabolism; D008861:Microsomes / Q000201:enzymology*; D010743:Phospholipids / Q000378:metabolism; D011188:Potassium / Q000378:metabolism; D011817:Rabbits; D012964:Sodium / Q000378:metabolism; D013347:Subcellular Fractions / Q000201:enzymology
D003902:Detergents; D010743:Phospholipids; D003840:Deoxycholic Acid; D012964:Sodium; D000251:Adenosine Triphosphatases; D011188:Potassium