Skip to main content
SpringerLink
Log in

Different sensitivity of ATP + Mg + Na (I) and Pi + Mg (II) dependent types of ouabain binding to phospholipase A2

  • Articles
  • Published:
The Journal of Membrane Biology Aims and scope Submit manuscript

Summary

The effect of phospholipase A2 and of related agents on ouabain binding and Na,K-ATPase activity were studied in intact and detergent-treated membrane preparations of rat brain cortex and pig kidney medulla. It was found that phospholipase A2 (PLA2) may distinguish or dissociate ouabain binding complexes I (ATP+Mg+Na) and II (Pi+Mg), stimulating the former and inhibiting the latter. Procedures which break the permeability barriers of vesicular membrane preparations, such as repeated freezing-thawing, sonication or hypoosmotic shock failed to mimic the effect of PLA2, indicating that it was not acting primarily by opening the inside-out oriented vesicles. The detergent digitonin exhibited similar effects on ouabain binding in both ATP+Mg+Na and Pi+Mg media. Other detergents were ineffective.

The ability of PLA2 to distinguish between ouabain binding type I and II can be manifested even in SDS-treated, purified preparations of Na,K-ATPase. The number of ATP+Mg+Na-dependent sites is unchanged, while the Pi+Mg-dependent sites are decreased in number in a manner similar to that seen in original membranes. This inhibition is completely lost in the reconstituted Na,K-ATPase system, where the ATP- as well as Pi-oriented ouabain sites are inhibited by PLA2.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Abeywardena, M.Y., Charnock, J.S. 1983. Modulation of cardiac glycoside inhibition of Na, K-ATPase by membranes: Difference between species.Biochim. Biophys. Acta 729:75–84

    PubMed  Google Scholar 

  • Abeywardena, M.Y., McMurichie, E.J., Russel, G.R., Charnock, J.S. 1984. Species variation in the ouabain sensitivity of cardiac Na, K-ATPase: A possible role for membrane lipids.Biochem. Pharmacol. 33:3649–3654

    PubMed  Google Scholar 

  • Albers, R.W., Koval, G.J., Siegel, G.J. 1968. Studies on the interaction of ouabain and other cardioactive steroids with sodium-potassium activated adenosine triphosphatase.Mol. Pharmacol. 4:324–336

    PubMed  Google Scholar 

  • Bodemann, H.H., Hoffman, J.F. 1976a. Side-dependent effects of internal versus external Na and K on ouabain binding to reconstituted human red blood cells ghosts.J. Gen. Physiol. 67:497–525

    PubMed  Google Scholar 

  • Bodemann, H.H., Hoffman, J.F. 1976b. Comparison of the side-dependent effects of Na and K on orthophosphate-, UTP- and ATP-promoted ouabain binding to reconstituted human red blood cell ghosts.J. Gen. Physiol. 67:527–545

    PubMed  Google Scholar 

  • Cantley, L.C. 1980. Structure and mechanism of the Na, K-ATPase.Curr. Topics Bioenerg. 11:201–237

    Google Scholar 

  • Charnock, J.S., Simonson, L.P., Almeida, A.F. 1977. Variation in sensitivity of the cardiac glycoside receptor characteristics of Na, K-ATPase to lipolysis and temperature.Biochim. Biophys. Acta 465:77–92

    PubMed  Google Scholar 

  • Cressie, L.C., Keightley, O.D. 1981. Analysing data from hormone-receptor assays.Biometrics 37:235–249

    PubMed  Google Scholar 

  • De Robertis, E., DeIraldi, A.P., DeLorez, G.R., Salganicoff, A.L. 1962. Cholinergic and noncholinergic nerve endings in the rat brain.J. Neurochem. 9:23–35

    PubMed  Google Scholar 

  • Erdman, E., Schoner, W. 1973. Ouabain-receptor interactions in Na,K-ATPase preparations: III. On the stability of the ouabain receptor against physical treatment, hydrolases and SH reagents.Biochim. Biophys. Acta 330:316–324

    PubMed  Google Scholar 

  • Forbush, B. III. 1982. Characterisation of right-side-out membrane vesicles rich in Na,K-ATPase and isolated from dog kidney medulla.J. Biol. Chem. 257:12678–12684

    PubMed  Google Scholar 

  • Fortes, P.A.G. 1977. Anthroylouabain A: A specific fluorescent probe for the cardiac glycoside receptor on Na,K-ATPase.Biochemistry 16:531–540

    PubMed  Google Scholar 

  • Garrahan, P.J., Horestein, A., Rega, A.F. 1977. The interaction of ligands with the Na,K-ATPase during Na,K-ATPase activity.In: Na,K-ATPase: Structure and kinetics. J.C. Skou and J.G. Norby, editors. pp. 261–274. Academic, London

    Google Scholar 

  • Goldmann, S.S., Albers, R.W. 1973. Sodium-potassium activated adenosine triphosphatase: IX. The role of phospholipids.J. Biol. Chem. 248:867–874

    PubMed  Google Scholar 

  • Goodman, S.L., Wheeler, K.P. 1978. Ouabain binding to phospholipid dependent adenosine triphosphatase.Biochem. J. 169:313–320

    PubMed  Google Scholar 

  • Jones, L.R., Maddock, S.W., Besh, H.R. 1980. Unmasking effect of alamethicin on beta-adrenergic receptor-coupled adenylate cyclase and cAMP-dependent protein kinase activities of cardiac sarcolemmal vesicles.J. Biol. Chem. 255:9971–9980

    Google Scholar 

  • Jorgensen, P.L. 1975. Isolation and characterization of the components of the sodium pump.Q. Rev. Biophys. 7:239–274

    Google Scholar 

  • Jorgensen, P.L. 1974. Purification and characterization of Na,K-ATPase: III. Purification from the outer medulla of mammalian kidney after selective removal of membrane components by SDS.Biochim. Biophys. Acta 356:36–52

    PubMed  Google Scholar 

  • Jorgensen, P.L., Skou, J.C. 1971. Purification and characterization of Na,K-ATPase: I. Influence of detergents on the activity of Na,K-ATPase in preparations from the outer medulla of rabbit kidney.Biochim. Biophys. Acta 233:366–380

    PubMed  Google Scholar 

  • Karlish, S.J.D., Pick, U. 1981. Sidedness of the effects of sodium and potassium ions on the conformational state of the sodium-potassium pump.J. Physiol. (London) 312:505–529

    Google Scholar 

  • Kimmelberg, H.K., Papahadjopoulos, D., 1974. Effects of phospholipid acyl chain fluidity, phase transitions and cholesterol on Na,K-stimulated triphosphatase.J. Biol. Chem. 249:1071–1080

    Google Scholar 

  • Lane, L.K., Anner, B.M., Wallick, E.T., Ray, M.V., Schwartz, A. 1978. Effect of phospholipase treatment on the partial reactions of and ouabain binding to a purified sodium and potassium activated adenosine triphosphatase.Biochem. Pharmacol. 27:225–231

    PubMed  Google Scholar 

  • Lee, J.A., Fortes, P.A.G. 1985. Anthroylouabain binding to different forms of Na,K-ATPase.In: The Sodium Pump. I. M. Glynn and J. C. Ellory, editors. pp. 277–282. Company of Biologists, Cambridge, England

    Google Scholar 

  • Lindenmayer, G.E., Schwartz, A. 1973. Nature of the transport adenosine triphosphatase digitalis complex: IV. Evidence that sodium-potassium competition modulates the rate of ouabain interaction with Na,K-adenosine-triphosphatase during enzyme catalysis.J. Biol. Chem. 248:1291–1300

    PubMed  Google Scholar 

  • Lin-Shiau, S.Y., Chen, C.C. 1982. Effects of beta-bungarotoxin and phospholipase A2 from Naja Naja snake venom on ATPase activities of synaptic membranes from rat cerebral cortex.Toxicon 20:409–418

    PubMed  Google Scholar 

  • Matsuda, T., Iwata, H. 1986. Phospholipid composition of cardiac Na,K-ATPase from various species.Experientia 42:405–407

    Google Scholar 

  • Moczydlowski, E.G., Fortes, P.A.G. 1980. Kinetics of cardiac glycoside binding to Na,K-adenosine triphosphatase studied with a fluorescence derivative of ouabain.Biochemistry 19:969–977

    PubMed  Google Scholar 

  • Pullman, M.E., Penefsky, H.S., Datta, A., Racker, E. 1960. Partial resolution of the enzymes catalyzing oxidative phosphorylation: I. Purification and properties of soluble dinitrophenol stimulated adenosine triphosphatase.J. Biol. Chem. 235:3322–3329

    PubMed  Google Scholar 

  • Roelofsen, B. 1981. The (non) specificity in the lipid-requirement of calcium and sodium plus potassium-transporting adenosine triphosphatases.Life Sci. 29:2235–2247

    PubMed  Google Scholar 

  • Roelofsen, B., Van Deenen, L.L.M., 1973. Lipid requirement of membrane bound ATPase: Studies on human erythrocyte ghosts.Eur. J. Biochem. 40:245–257

    Google Scholar 

  • Scatchard, G. 1949. The attraction of proteins for small molecules and ions.Ann. N.Y. Acad. Sci. 51:660–672

    Google Scholar 

  • Schmalzing, G., Kutschera, P. 1982. Modulation of ATPase activities of human erythrocyte membranes by free fatty acids or phospholipase A2.J. Membrane Biol. 69:65–76

    Google Scholar 

  • Schull, G.E., Schwartz, A., Lindgrel, J.B., 1985. Amino acid sequence of the catalytical subunit of the Na,K-ATPase deduced from the complementary DNA.Nature (London) 316:691–695

    Google Scholar 

  • Stahl, W.L. 1986. The Na,K-ATPase of the nervous tissue.Neurochem. Int. 8:449–476

    Google Scholar 

  • Svoboda, P., Mosinger, B. 1981. Catecholamines and the rat brain microsomal Na,K-ATPase: I. Protection against lipoperoxidative damage.Biochem. Pharmacol. 30:427–432

    PubMed  Google Scholar 

  • Sweadner, K.J. 1978. Purification from rat brain of an intrisic membrane protein fraction enriched in Na,K-ATPase.Biochim. Biophys. Acta 508:486–499

    PubMed  Google Scholar 

  • Sweadner, K.J. 1979. Two molecular forms of Na,K-stimulated ATPase in brain. Separation and difference in affinity for strophantidin.J. Biol. Chem. 254:6060–6067

    Google Scholar 

  • Sweadner, K.J. 1985. Enzymatic properties of separated isoenzymes of the Na,K-ATPase: Substrate affinities, kinetic cooperativity and ion transport stoichimetry.J. Biol. Chem. 260:11508–11513

    PubMed  Google Scholar 

  • Taniguchi, K., Iida, S. 1973. The role of phospholipids in the binding of ouabain to sodium and potassium dependent adenosine triphosphatase.Mol. Pharmacol. 9:350–359

    PubMed  Google Scholar 

  • Taussky, H.H., Shorr, E. 1953. A microcolorimetric method for the determination of inorganic phosphorus.J. Biol. Chem. 202:675–685

    PubMed  Google Scholar 

  • Walter, H. 1975. Tightness and orientation of vesicles from guinea-pig kidney estimated from reaction of adenosine triphosphatase dependent on sodium and potassium ions.Eur. J. Biochem. 58:595–601

    PubMed  Google Scholar 

  • Walter, H. 1979. Permeability of plasma membrane vesicles to ouabain and Mg2+ as a factor determining the rate of binding of ouabain to Na,K-ATPase.Z. Naturforsch. 34:1224–1231

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institute of Physiology, Czechoslovak Academy of Sciences, Videnska 1083, 4, Praha, Czechoslovakia

    Petr Svoboda, Evzen Amler & Jan Teisinger

Authors
  1. Petr Svoboda
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Evzen Amler
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jan Teisinger
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Svoboda, P., Amler, E. & Teisinger, J. Different sensitivity of ATP + Mg + Na (I) and Pi + Mg (II) dependent types of ouabain binding to phospholipase A2 . J. Membrain Biol. 104, 211–221 (1988). https://doi.org/10.1007/BF01872323

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01872323

Key Words

Search

Navigation