Skip to main content
NCBI home page
Biophysical Journal logoLink to Biophysical Journal
. 1999 Jul;77(1):267–281. doi: 10.1016/S0006-3495(99)76888-8

Hofmeister effects of anions on the kinetics of partial reactions of the Na+,K+-ATPase.

C Ganea 1, A Babes 1, C Lüpfert 1, E Grell 1, K Fendler 1, R J Clarke 1
PMCID: PMC1300328  PMID: 10388756

Abstract

The effects of lyotropic anions, particularly perchlorate, on the kinetics of partial reactions of the Na+,K+-ATPase from pig kidney were investigated by two different kinetic techniques: stopped flow in combination with the fluorescent label RH421 and a stationary electrical relaxation technique. It was found that 130 mM NaClO4 caused an increase in the Kd values of both the high- and low-affinity ATP-binding sites, from values of 7.0 (+/- 0.6) microM and 143 (+/- 17) microM in 130 mM NaCl solution to values of 42 (+/- 3) microM and 660 (+/- 100) microM in 130 mM NaClO4 (pH 7.4, 24 degrees C). The half-saturating concentration of the Na+-binding sites on the E1 conformation was found to decrease from 8-10 mM in NaCl to 2.5-3.5 mM in NaClO4 solution. The rate of equilibration of the reaction, E1P(Na+)3 left arrow over right arrow E2P + 3Na+, decreased from 393 (+/- 51) s-1 in NaCl solution to 114 (+/- 15) s-1 in NaClO4. This decrease is attributed predominantly to an inhibition of the E1P(Na+)3 --> E2P(Na+)3 transition. The effects can be explained in terms of electrostatic interactions due to perchlorate binding within the membrane and/or protein matrix of the Na+,K+-ATPase membrane fragments and alteration of the local electric field strength experienced by the protein. The kinetic results obtained support the conclusion that the conformational transition E1P(Na+)3 --> E2P(Na+)3 is a major charge translocating step of the pump cycle.

Full Text

The Full Text of this article is available as a PDF (156.1 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Cacace M. G., Landau E. M., Ramsden J. J. The Hofmeister series: salt and solvent effects on interfacial phenomena. Q Rev Biophys. 1997 Aug;30(3):241–277. doi: 10.1017/s0033583597003363. [DOI] [PubMed] [Google Scholar]
  2. Clarke R. J. Effect of lipid structure on the dipole potential of phosphatidylcholine bilayers. Biochim Biophys Acta. 1997 Jul 25;1327(2):269–278. doi: 10.1016/s0005-2736(97)00075-8. [DOI] [PubMed] [Google Scholar]
  3. Clarke R. J., Kane D. J., Apell H. J., Roudna M., Bamberg E. Kinetics of Na(+)-dependent conformational changes of rabbit kidney Na+,K(+)-ATPase. Biophys J. 1998 Sep;75(3):1340–1353. doi: 10.1016/S0006-3495(98)74052-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Clarke R. J., Lüpfert C. Influence of anions and cations on the dipole potential of phosphatidylcholine vesicles: a basis for the Hofmeister effect. Biophys J. 1999 May;76(5):2614–2624. doi: 10.1016/S0006-3495(99)77414-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Collins K. D., Washabaugh M. W. The Hofmeister effect and the behaviour of water at interfaces. Q Rev Biophys. 1985 Nov;18(4):323–422. doi: 10.1017/s0033583500005369. [DOI] [PubMed] [Google Scholar]
  6. Cornelius F., Fedosova N. U., Klodos I. E2P phosphoforms of Na,K-ATPase. II. Interaction of substrate and cation-binding sites in Pi phosphorylation of Na,K-ATPase. Biochemistry. 1998 Nov 24;37(47):16686–16696. doi: 10.1021/bi981571v. [DOI] [PubMed] [Google Scholar]
  7. Cornelius F. Hydrophobic ion interaction on Na+ activation and dephosphorylation of reconstituted Na+,K(+)-ATPase. Biochim Biophys Acta. 1995 May 4;1235(2):183–196. doi: 10.1016/0005-2736(95)80004-y. [DOI] [PubMed] [Google Scholar]
  8. Dani J. A., Sanchez J. A., Hille B. Lyotropic anions. Na channel gating and Ca electrode response. J Gen Physiol. 1983 Feb;81(2):255–281. doi: 10.1085/jgp.81.2.255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fendler K., Grell E., Haubs M., Bamberg E. Pump currents generated by the purified Na+K+-ATPase from kidney on black lipid membranes. EMBO J. 1985 Dec 1;4(12):3079–3085. doi: 10.1002/j.1460-2075.1985.tb04048.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fendler K., Jaruschewski S., Hobbs A., Albers W., Froehlich J. P. Pre-steady-state charge translocation in NaK-ATPase from eel electric organ. J Gen Physiol. 1993 Oct;102(4):631–666. doi: 10.1085/jgp.102.4.631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Flewelling R. F., Hubbell W. L. The membrane dipole potential in a total membrane potential model. Applications to hydrophobic ion interactions with membranes. Biophys J. 1986 Feb;49(2):541–552. doi: 10.1016/S0006-3495(86)83664-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Frank J., Zouni A., van Hoek A., Visser A. J., Clarke R. J. Interaction of the fluorescent probe RH421 with ribulose-1,5-bisphosphate carboxylase/oxygenase and with Na+,K(+)-ATPase membrane fragments. Biochim Biophys Acta. 1996 Apr 3;1280(1):51–64. doi: 10.1016/0005-2736(95)00277-4. [DOI] [PubMed] [Google Scholar]
  13. Friedrich T., Nagel G. Comparison of Na+/K(+)-ATPase pump currents activated by ATP concentration or voltage jumps. Biophys J. 1997 Jul;73(1):186–194. doi: 10.1016/S0006-3495(97)78059-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. HODGKIN A. L., HOROWICZ P. The effect of nitrate and other anions on the mechanical response of single muscle fibres. J Physiol. 1960 Sep;153:404–412. doi: 10.1113/jphysiol.1960.sp006542. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hobbs A. S., Albers R. W., Froehlich J. P. Potassium-induced changes in phosphorylation and dephosphorylation of (Na+ + K+)-ATPase observed in the transient state. J Biol Chem. 1980 Apr 25;255(8):3395–3402. [PubMed] [Google Scholar]
  16. Jorgensen P. L. Isolation of (Na+ plus K+)-ATPase. Methods Enzymol. 1974;32:277–290. [PubMed] [Google Scholar]
  17. Jorgensen P. L. Purification and characterization of (Na+ plus K+ )-ATPase. 3. Purification from the outer medulla of mammalian kidney after selective removal of membrane components by sodium dodecylsulphate. Biochim Biophys Acta. 1974 Jul 12;356(1):36–52. doi: 10.1016/0005-2736(74)90292-2. [DOI] [PubMed] [Google Scholar]
  18. Jørgensen P. L., Andersen J. P. Structural basis for E1-E2 conformational transitions in Na,K-pump and Ca-pump proteins. J Membr Biol. 1988 Jul;103(2):95–120. doi: 10.1007/BF01870942. [DOI] [PubMed] [Google Scholar]
  19. Kane D. J., Fendler K., Grell E., Bamberg E., Taniguchi K., Froehlich J. P., Clarke R. J. Stopped-flow kinetic investigations of conformational changes of pig kidney Na+,K+-ATPase. Biochemistry. 1997 Oct 28;36(43):13406–13420. doi: 10.1021/bi970598w. [DOI] [PubMed] [Google Scholar]
  20. Kane D. J., Grell E., Bamberg E., Clarke R. J. Dephosphorylation kinetics of pig kidney Na+,K+-ATPase. Biochemistry. 1998 Mar 31;37(13):4581–4591. doi: 10.1021/bi972813e. [DOI] [PubMed] [Google Scholar]
  21. Klodos I., Fedosova N. U., Plesner L. Influence of intramembrane electric charge on Na,K-ATPase. J Biol Chem. 1995 Mar 3;270(9):4244–4254. doi: 10.1074/jbc.270.9.4244. [DOI] [PubMed] [Google Scholar]
  22. Klodos I., Post R. L., Forbush B., 3rd Kinetic heterogeneity of phosphoenzyme of Na,K-ATPase modeled by unmixed lipid phases. Competence of the phosphointermediate. J Biol Chem. 1994 Jan 21;269(3):1734–1743. [PubMed] [Google Scholar]
  23. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  24. Liberman E. A., Topaly V. P. Pronitsaemost' bimolekuliarnykh fosfolipidnykh membran dlia zhirorastvorimykh ionov. Biofizika. 1969 May-Jun;14(3):452–461. [PubMed] [Google Scholar]
  25. Lu C. C., Kabakov A., Markin V. S., Mager S., Frazier G. A., Hilgemann D. W. Membrane transport mechanisms probed by capacitance measurements with megahertz voltage clamp. Proc Natl Acad Sci U S A. 1995 Nov 21;92(24):11220–11224. doi: 10.1073/pnas.92.24.11220. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. McLaughlin S., Bruder A., Chen S., Moser C. Chaotropic anions and the surface potential of bilayer membranes. Biochim Biophys Acta. 1975 Jun 25;394(2):304–313. doi: 10.1016/0005-2736(75)90267-9. [DOI] [PubMed] [Google Scholar]
  27. Nørby J. G., Esmann M. The effect of ionic strength and specific anions on substrate binding and hydrolytic activities of Na,K-ATPase. J Gen Physiol. 1997 May;109(5):555–570. doi: 10.1085/jgp.109.5.555. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Peters W. H., Swarts H. G., de Pont J. J., Schuurmans Stekhoven F. M., Bonting S. L. (Na/ + K+)ATPase has one functioning phosphorylation site per alpha subunit. Nature. 1981 Mar 26;290(5804):338–339. doi: 10.1038/290338a0. [DOI] [PubMed] [Google Scholar]
  29. Post R. L., Klodos I. Interpretation of extraordinary kinetics of Na(+)-K(+)-ATPase by a phase change. Am J Physiol. 1996 Nov;271(5 Pt 1):C1415–C1423. doi: 10.1152/ajpcell.1996.271.5.C1415. [DOI] [PubMed] [Google Scholar]
  30. Post R. L., Suzuki K. A Hofmeister effect on the phosphoenzyme of Na,K-ATPase. Soc Gen Physiol Ser. 1991;46:201–209. [PubMed] [Google Scholar]
  31. Pratap P. R., Robinson J. D. Rapid kinetic analyses of the Na+/K(+)-ATPase distinguish among different criteria for conformational change. Biochim Biophys Acta. 1993 Sep 5;1151(1):89–98. doi: 10.1016/0005-2736(93)90075-b. [DOI] [PubMed] [Google Scholar]
  32. Rychkov G. Y., Pusch M., Roberts M. L., Jentsch T. J., Bretag A. H. Permeation and block of the skeletal muscle chloride channel, ClC-1, by foreign anions. J Gen Physiol. 1998 May;111(5):653–665. doi: 10.1085/jgp.111.5.653. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Sokolov V. S., Stukolov S. M., Darmostuk A. S., Apell H. J. Influence of sodium concentration on changes of membrane capacitance associated with the electrogenic ion transport by the Na,K-ATPase. Eur Biophys J. 1998;27(6):605–617. doi: 10.1007/s002490050172. [DOI] [PubMed] [Google Scholar]
  34. Sokolov V. S., Stukolov S. M., Gevondyan N. M., Apell H. J. Study of electrogenic transport of sodium ions inside the Na,K-ATPase by means of membrane capacitance measurements. Ann N Y Acad Sci. 1997 Nov 3;834:364–366. doi: 10.1111/j.1749-6632.1997.tb52272.x. [DOI] [PubMed] [Google Scholar]
  35. Stürmer W., Bühler R., Apell H. J., Läuger P. Charge translocation by the Na,K-pump: II. Ion binding and release at the extracellular face. J Membr Biol. 1991 Apr;121(2):163–176. doi: 10.1007/BF01870530. [DOI] [PubMed] [Google Scholar]
  36. Suzuki K., Post R. L. Equilibrium of phosphointermediates of sodium and potassium ion transport adenosine triphosphatase: action of sodium ion and Hofmeister effect. J Gen Physiol. 1997 May;109(5):537–554. doi: 10.1085/jgp.109.5.537. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Tatulian S. A. Binding of alkaline-earth metal cations and some anions to phosphatidylcholine liposomes. Eur J Biochem. 1987 Dec 30;170(1-2):413–420. doi: 10.1111/j.1432-1033.1987.tb13715.x. [DOI] [PubMed] [Google Scholar]
  38. Tatulian S. A. Effect of lipid phase transition on the binding of anions to dimyristoylphosphatidylcholine liposomes. Biochim Biophys Acta. 1983 Dec 21;736(2):189–195. doi: 10.1016/0005-2736(83)90283-3. [DOI] [PubMed] [Google Scholar]
  39. Wuddel I., Apell H. J. Electrogenicity of the sodium transport pathway in the Na,K-ATPase probed by charge-pulse experiments. Biophys J. 1995 Sep;69(3):909–921. doi: 10.1016/S0006-3495(95)79965-9. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Biophysical Journal are provided here courtesy of The Biophysical Society

RESOURCES