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Abstract:

This work presents a detailed kinetic study that shows the coupling between the E2→E1 transition and Rb+ deocclusion stimulated by Na + in pig-kidney purified Na,K-ATPase. Using rapid mixing techniques, we measured in parallel experiments the decrease in concentration of occluded Rb+ and the increase in eosin fluorescence (the formation of E1) as a function of time. The E2→E1 transition and Rb+ deocclusion are described by the sum of two exponential functions with equal amplitudes, whose rate coefficients decreased with increasing [Rb+]. The rate coefficient values of the E2→E1 transition were very similar to those of Rb+-deocclusion, indicating that both processes are simultaneous. Our results suggest that, when ATP is absent, the mechanism of Na +-stimulated Rb+ deocclusion would require the release of at least one Rb+ ion through the extracellular access prior to the E2→E1 transition. Using vanadate to stabilize E2, we measured occluded Rb+ in equilibrium conditions. Results show that, while Mg 2 + decreases the affinity for Rb+, addition of vanadate offsets this effect, increasing the affinity for Rb+. In transient experiments, we investigated the exchange of Rb+ between the E2-vanadate complex and the medium. Results show that, in the absence of ATP, vanadate prevents the E2→E1 transition caused by Na+ without significantly affecting the rate of Rb+ deocclusion. On the other hand, we found the first evidence of a very low rate of Rb+ occlusion in the enzyme-vanadate complex, suggesting that this complex would require a change to an open conformation in order to bind and occlude Rb+. © 2012 Elsevier B.V. All rights reserved.

Registro:

Documento: Artículo
Título:E2→E1 transition and Rb+ release induced by Na+ in the Na+/K+-ATPase. Vanadate as a tool to investigate the interaction between Rb+ and E2
Autor:Montes, M.R.; Monti, J.L.E.; Rossi, R.C.
Filiación:Instituto de Química Y Fisicoquímica Biológicas, Departamento de Química Biológica, Universidad de Buenos Aires, Junín 956, 1113 Buenos Aires, Argentina
Agencia Nacional de Promoción Científica Y Tecnológica, Argentina
Palabras clave:Conformational transition; Na+/K+-ATPase; Rb +-deocclusion; Vanadate; adenosine triphosphatase (potassium sodium); eosin; magnesium ion; rubidium ion; sodium ion; vanadic acid; amplitude modulator; animal tissue; article; binding affinity; conformational transition; enzyme inhibitor interaction; equilibrium constant; fluorescence analysis; nonhuman; phase transition; priority journal; Adenosine Triphosphate; Animals; Biophysics; Eosine Yellowish-(YS); Kidney; Kinetics; Magnesium; Models, Biological; Protein Binding; Protein Conformation; Rubidium; Sodium-Potassium-Exchanging ATPase; Swine; Time Factors; Vanadates; Suidae
Año:2012
Volumen:1818
Número:9
Página de inicio:2087
Página de fin:2093
DOI: http://dx.doi.org/10.1016/j.bbamem.2012.04.001
Título revista:Biochimica et Biophysica Acta - Biomembranes
Título revista abreviado:Biochim. Biophys. Acta Biomembr.
ISSN:00052736
CODEN:BBBMB
CAS:eosin, 17372-87-1, 51395-88-1, 548-26-5; magnesium ion, 22537-22-0; rubidium ion, 22537-38-8; sodium ion, 17341-25-2; vanadic acid, 12260-63-8, 13981-20-9, 37353-31-4; Adenosine Triphosphate, 56-65-5; Eosine Yellowish-(YS), 17372-87-1; Magnesium, 7439-95-4; Rubidium, 7440-17-7; Sodium-Potassium-Exchanging ATPase, 3.6.3.9; Vanadates
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00052736_v1818_n9_p2087_Montes

Referencias:

  • Glynn, I.M., All hands to the sodium pump (1993) J. Physiol., 462, pp. 1-30
  • Glynn, I.M., Hara, Y., Richards, D.E., Steinberg, M., Comparison of rates of cation release and of conformational change in dog kidney Na,K-ATPase (1987) Journal of Physiology, VOL. 383, pp. 477-485
  • Esmann, M., Influence of Na+on conformational states in membrane-bound renal Na,K-ATPase (1994) Biochemistry, 33, pp. 8558-8565
  • Karlish, S.J., Beaugé, L.A., Glynn, I.M., Vanadate inhibits (Na+/K+)ATPase by blocking a conformational change of the unphosphorylated form (1979) Nature, 282 (5736), pp. 333-335
  • Glynn, I.M., Richards, D.E., Occlusion of rubidium ions by the sodium-potassium pump: Its implications for the mechanism of potassium transport (1982) Journal of Physiology, VOL. 330, pp. 17-43
  • Montes, M.R., Spiaggi, A.J., Monti, J.L., Cornelius, F., Olesen, C., Garrahan, P.J., Rossi, R.C., Rb+ occlusion stabilized by vanadate in gastric H+/K+-ATPase at 25°C (2011) Biochim. Biophys. Acta, 1808, pp. 316-322
  • Abe, K., Unique structures and conserved conformational changes found in gastric (1723) H,K-ATPase, Na,K-ATPase and related P-ATPases: Structure, biology and medicine 13th International P-type ATPase Meeting, 2011
  • Rabon, E.C., Smillie, K., Seru, V., Rabon, R., Rubidium occlusion within tryptic peptides of the H,K-ATPase (1993) Journal of Biological Chemistry, 268 (11), pp. 8012-8018
  • Clausen, J.D., McIntosh, D.B., Woolley, D.G., Andersen, J.P., Modulatory ATP binding affinity in intermediate states of E2P dephosphorylation of sarcoplasmic reticulum Ca2+-ATPase (2011) J. Biol. Chem., 286, pp. 11792-11802
  • Forbush, B., Rapid release of 42K or 86Rb from two distinct transport sites on the Na, K-pump in the presence of Pi or vanadate (1987) J. Biol. Chem., 262 (23), pp. 11116-11127
  • Jensen, J., Norby, J.G., Ottolenghi, P., Binding of sodium and potassium to the sodium pump of pig kidney evaluated from nucleotide-binding behaviour (1984) Journal of Physiology, VOL. 346, pp. 219-241
  • Rossi, R.C., Kaufman, S.B., Gonzalez, L.R.M., Norby, J.G., Garrahan, P.J., An attachment for nondestructive, fast quenching of samples in rapid- mixing experiments (1999) Analytical Biochemistry, 270 (2), pp. 276-285. , DOI 10.1006/abio.1999.4094
  • Skou, J.C., Esmann, M., Eosin, a fluorescent probe of ATP binding to the (Na+-K +)- ATPase (1981) Biochim. Biophys. Acta, 647, pp. 232-240
  • Montes, M.R., González-Lebrero, R.M., Garrahan, P.J., Rossi, R.C., Eosin fluorescence changes during Rb+ occlusion in the Na +/K+-ATPase (2006) Biochemistry, 45, pp. 13093-13100
  • González-Lebrero, R.M., Kaufman, S.B., Garrahan, P.J., Rossi, R.C., The occlusion of Rb+in the Na,K-ATPase II (2002) J. Biol. Chem., 277, pp. 5922-5928
  • Skou, J.C., Esmann, M., The effects of Na+and K+on the conformational transitions of (Na++K+)-ATPase (1983) Biochim. Biophys. Acta, 746, pp. 101-113
  • Fedosova, N.U., Esmann, M., Nucleotide-Binding Kinetics of Na,K-ATPase: Cation Dependence (2004) Biochemistry, 43 (14), pp. 4212-4218. , DOI 10.1021/bi035707n
  • Karlish, S.J., Characterization of conformational changes in (Na,K) ATPase labeled with fluorescein at the active site (1980) J. Bioenerg. Biomembr., 3-4, pp. 111-136
  • Faller, L.D., Scheiner-Bobis, R.A., Farley, R.A., Temperature dependence of the rates of conformational changes reported by fluorescein 5′-isothiocyanate (1991) Biochemistry, 30 (14), pp. 3503-3510
  • Humprey, P.A., Lupfert, C., Apell, H., Cornelius, F., Clarke, R., Mechanism of the rate-determining step of the Na+,K +-ATPase pump cycle (2002) Biochemistry, 41, pp. 9496-9507
  • Forbush III, B., Rapid release of42K and86Rb from an occluded state of the Na,K-pump in the presence of ATP or ADP (1987) J. Biol. Chem., 262, pp. 11104-11115
  • Forbush III, B., Occluded ions and Na,K-ATPase (1988) Progress in Clinical and Biological Research, 258. , Alan R. Liss, Inc., New York
  • Cantley, L.C., Cantley, L.G., Josephson, L., A characterization of vanadate interactions with the (Na,K)-ATPase (1984) J. Biol. Chem., 253 (20), pp. 7361-7368
  • Post, R.L., Toda, G., Rogers, F.N., Phosphorylation by inorganic phosphate of sodium plus potassium ion transport adenosine triphosphatase (1975) J. Biol. Chem., 250, pp. 691-701
  • Glynn, I.M., Karlish, S.J., Occluded cations in active transport (1990) Annu. Rev. Biochem., 59, pp. 171-205

Citas:

---------- APA ----------
Montes, M.R., Monti, J.L.E. & Rossi, R.C. (2012) . E2→E1 transition and Rb+ release induced by Na+ in the Na+/K+-ATPase. Vanadate as a tool to investigate the interaction between Rb+ and E2. Biochimica et Biophysica Acta - Biomembranes, 1818(9), 2087-2093.
http://dx.doi.org/10.1016/j.bbamem.2012.04.001
---------- CHICAGO ----------
Montes, M.R., Monti, J.L.E., Rossi, R.C. "E2→E1 transition and Rb+ release induced by Na+ in the Na+/K+-ATPase. Vanadate as a tool to investigate the interaction between Rb+ and E2" . Biochimica et Biophysica Acta - Biomembranes 1818, no. 9 (2012) : 2087-2093.
http://dx.doi.org/10.1016/j.bbamem.2012.04.001
---------- MLA ----------
Montes, M.R., Monti, J.L.E., Rossi, R.C. "E2→E1 transition and Rb+ release induced by Na+ in the Na+/K+-ATPase. Vanadate as a tool to investigate the interaction between Rb+ and E2" . Biochimica et Biophysica Acta - Biomembranes, vol. 1818, no. 9, 2012, pp. 2087-2093.
http://dx.doi.org/10.1016/j.bbamem.2012.04.001
---------- VANCOUVER ----------
Montes, M.R., Monti, J.L.E., Rossi, R.C. E2→E1 transition and Rb+ release induced by Na+ in the Na+/K+-ATPase. Vanadate as a tool to investigate the interaction between Rb+ and E2. Biochim. Biophys. Acta Biomembr. 2012;1818(9):2087-2093.
http://dx.doi.org/10.1016/j.bbamem.2012.04.001