Article: Reconstitution of the sodium pump from protein and phosphatidylserine: features of ouabain binding

1. A study has been made of ATP splitting and ouabain binding to the sodium pump reconstituted from protein and phosphatidylserine.2. Ouabain was bound to protein alone, but when phosphatidylserine was added, binding was increased threefold. The stimulation resembled the course of activation of sodium-dependent ATPase activity.3. EGTA partly simulated the activation of ATPase by phosphatidylserine but did not enhance binding.4. The dissociation constant for the enzyme-ouabain complex was 3.5 x 10(-8)M. The turnover number (2,000 molecules of ATP per minute) and the number of receptor sites (3.8 x 10(13) per mg protein) were calculated.5. The results provide further evidence of the involvement of phosphatidylserine in the action of the sodium pump.

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References

1.

Hendrickson H, Fullington J . Stabilities of metal complexes of phospholipids: Ca(II), Mg(II), and Ni(II) complexes of phosphatidylserine and triphosphoinositide. Biochemistry. 1965; 4(8):1599-605. DOI: 10.1021/bi00884a021. View

2.

Tanaka R, Strickland K . Role of phospholipid in the activation of Na+, Ka+-activated adenosine triphosphatase of beef brain. Arch Biochem Biophys. 1965; 111(3):583-92. DOI: 10.1016/0003-9861(65)90239-0. View

3.

Fenster L, COPENHAVER Jr J . Phosphatidyl serine requirement of (Na+-K+)-activated adenosine triphosphatase from rat kidney and brain. Biochim Biophys Acta. 1967; 137(2):406-8. DOI: 10.1016/0005-2760(67)90120-8. View

4.

Whittam R, WHEELER K . Transport across cell membranes. Annu Rev Physiol. 1970; 32:21-60. DOI: 10.1146/annurev.ph.32.030170.000321. View

5.

WHEELER K, Whittam R . The involvement of phosphatidylserine in adenosine triphosphatase activity of the sodium pump. J Physiol. 1970; 207(2):303-28. PMC: 1348708. DOI: 10.1113/jphysiol.1970.sp009063. View

6.

Hansen O . The relationship between G-strophanthin-binding capacity and ATPase activity in plasma-membrane fragments from ox brain. Biochim Biophys Acta. 1971; 233(1):122-32. DOI: 10.1016/0005-2736(71)90364-6. View

7.

Taniguchi K, Iida S . The binding of ouabain to Na + -K + -dependent ATPase treated with phospholipase. Biochim Biophys Acta. 1971; 233(3):831-3. DOI: 10.1016/0005-2736(71)90186-6. View

8.

Priestland R, Whittam R . The temperature dependence of activation by phosphatidylserine of the sodium pump adenosine triphosphatase. J Physiol. 1972; 220(2):353-61. PMC: 1331707. DOI: 10.1113/jphysiol.1972.sp009711. View

9.

Baker P, WILLIS J . Binding of the cardiac glycoside ouabain to intact cells. J Physiol. 1972; 224(2):441-62. PMC: 1331499. DOI: 10.1113/jphysiol.1972.sp009904. View

10.

LOWRY O, ROSEBROUGH N, FARR A, RANDALL R . Protein measurement with the Folin phenol reagent. J Biol Chem. 1951; 193(1):265-75. View

11.

HUMMEL J, Dreyer W . Measurement of protein-binding phenomena by gel filtration. Biochim Biophys Acta. 1962; 63:530-2. DOI: 10.1016/0006-3002(62)90124-5. View

Reconstitution of the Sodium Pump from Protein and Phosphatidylserine: Features of Ouabain Binding