Role of phospholipid in the intermediate steps of the sodium-plus-potassium ion-dependent adenosine triphosphatase reaction.
The phosphorylation and dephosphorylation steps of the (Na-++K-+)-dependent ATPase (adenosine triphosphatase) (EC 3.6.1.3) reaction have been compared in 'normal', lipid-depleted and 'restored' membrane ATPase preparations. Partial lipid depletion was achieved by a single extraction with Lubrol W, and 'restoration' by adding pure phosphatidylserine. Gamma-32-P-labelled ATP was used for phosphorylation. The main findings were as follows. (1) Partial lipid depletion decreased but did not prevent Na-+-dependent phosphorylation, although it virtually abolished both Na-+-dependent and (Na-++K-+)-dependent ATPase activities. (2) 'Restoration' with phosphatidylserine produced an increment in phosphorylation that was the same in the presence and absence of added Na-+. (3) K-+ decreased the extent of Na-+-dependent phosphorylation of the depleted enzyme without producing a corresponding release of Pi. (4) K-+ rapidly decreased the extent of phosphorylation of the 'restored' enzyme to near-background value, with a concomitant release of Pi. (5) Na-+-dependent ATP hydrolysis was not restored. (6) The turnover of the 'restored' enzyme seemed to be higher than that of the 'normal' enzyme. The reaction sequence is discussed in relation to these results and the fact that the depleted enzyme retained about 50% of K-+-dependent phosphatase activity.
D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000818:Animals; D002614:Chelating Agents; D003902:Detergents; D006868:Hydrolysis; D007668:Kidney / Q000201:enzymology; D007700:Kinetics; D008861:Microsomes / Q000201:enzymology; D010710:Phosphates / Q000378:metabolism; D010718:Phosphatidylserines; D010743:Phospholipids / Q000378:metabolism*; D011188:Potassium / Q000378:metabolism; D011817:Rabbits; D012964:Sodium / Q000378:metabolism; D013997:Time Factors
D002614:Chelating Agents; D003902:Detergents; D010710:Phosphates; D010718:Phosphatidylserines; D010743:Phospholipids; D012964:Sodium; D000251:Adenosine Triphosphatases; D011188:Potassium