Artículo

El editor solo permite decargar el artículo en su versión post-print desde el repositorio. Por favor, si usted posee dicha versión, enviela a
Consulte el artículo en la página del editor
Consulte la política de Acceso Abierto del editor

Abstract:

We used suspensions of partially purified Na+/K +-ATPase from pig kidney to compare the effects of Rb+, as a K+ congener, on the time course and on the equilibrium values of eosin fluorescence and of Rb+ occlusion. Both sets of data were collected under identical conditions in the same enzyme preparations. The incubation media lacked ATP so that all changes led to an equilibrium distribution between enzyme conformers with and without bound eosin and with and without bound or occluded Rb+. Results showed that as Rb+ concentration was increased, the equilibrium value of fluorescence decreased and occlusion increased along rectangular hyperbolas with similar half-maximal values. The time courses of attainment of equilibrium showed an initial phase which was so quick as to fall below the time resolution of our rapid-mixing apparatus. This phase was followed by the sum of at least two exponential functions of time. In the case of fluorescence the fast exponential term accounted for a larger fraction of the time course than in the case of occlusion. Comparison between experimental and simulated results suggests that fluorescence changes express a process that is coupled to Rb+ occlusion but that is completed before occlusion reaches equilibrium. © 2006 American Chemical Society.

Registro:

Documento: Artículo
Título:Eosin fluorescence changes during Rb+ occlusion in the Na +/K+-ATPase
Autor:Montes, M.R.; González-Lebrero, R.M.; Garrahan, P.J.; Rossi, R.C.
Filiación:Instituto de Química y Fisicoquímica Biológicas, Departamento de Química Biológica, Universidad de Buenos Aires, Junín 956, C1113AAD Buenos Aires, Argentina
Palabras clave:Adenosinetriphosphate; Data structures; Fluorescence; Positive ions; Rubidium; Sodium; Suspensions (fluids); Congeners; Eosin fluorescence; Hyperbolas; Occlusions; Enzymes; adenosine triphosphatase (potassium sodium); eosin; rubidium 86; article; enzyme activity; enzyme kinetics; enzyme purification; fluorescence; kidney; nonhuman; priority journal; swine; Algorithms; Animals; Eosine Yellowish-(YS); Fluorescence; Ion Transport; Kinetics; Models, Chemical; Na(+)-K(+)-Exchanging ATPase; Protein Binding; Rubidium; Swine; Sus scrofa
Año:2006
Volumen:45
Número:43
Página de inicio:13093
Página de fin:13100
DOI: http://dx.doi.org/10.1021/bi060778i
Título revista:Biochemistry
Título revista abreviado:Biochemistry
ISSN:00062960
CODEN:BICHA
CAS:eosin, 17372-87-1, 51395-88-1, 548-26-5; rubidium 86, 14932-53-7; Eosine Yellowish-(YS), 17372-87-1; Na(+)-K(+)-Exchanging ATPase, EC 3.6.1.37; Rubidium, 7440-17-7
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00062960_v45_n43_p13093_Montes

Referencias:

  • Skou, J.C., Esmann, M., The Na, K.-ATPase (1992) J. Bioenerg. Biomembr., 24, pp. 249-281
  • González-Lebrero, R.M., Kaufman, S.B., Montes, M.R., Nørby, J.G., Garrahan, P.J., Rossi, R.C., The occlusion of Rb+ in the Na+/K +-ATPase. I. The identity of occluded states formed by the physiological or the direct routes. Occlusion/deocclusion kinetics throuah the direct route (2002) J. Biol. Chem., 277, pp. 5910-5921
  • Jørgensen, P.L., Petersen, J., High-affinity 86Rb-binding and structural changes in the alpha-subunil of Na+.K+-ATPase as detected by tryplic digestion and fluorescence analysis (1982) Biochim. Biophys. Acta, 705, pp. 38-47
  • 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) J. Physiol., 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
  • Rossi, R.C., Kaufman, S.B., González-Lebrero, R.M., Nørby, J.G., Garrahan, P.J., An attachment for non-destructive, fast quenching of samples in rapid mixing experiments (1999) Anal. Biochem., 270, pp. 276-285
  • Skou, J.C., Esmann, M., Effect of magnesium ions on the high-affinity binding of eosin to the (Na+ + K+)-ATPase (1983) Biochim. Biophys. Acta, 727, pp. 101-107
  • Montes, M.R., González-Lebrero, R.M., Garrahan, P.J., Rossi, R.C., The interaction between the fluorescent probe eosin and the Na +/K+-ATPase studied through Rb+ occlusion (2004) Biochemistry, 43, pp. 2062-2069
  • Jensen, J., Nerby, J.G., Ottolenghi, P., Binding of sodium and potassium to the sodium pump of pig kidney evaluated from nucleotide-binding behaviour (1984) J. Physiol. (Lond.), 346, pp. 219-241
  • Lin, S., Faller, L.D., Ionic-strength dependence of the conformational change in the unphosphorylated sodium pump (2005) Biochemistry, 44, pp. 1482-1494
  • Nerby, J.G., Esmann, M., The effect of ionic strength and specific anions on substrate binding and hydrolytic activities of Na, K.-ATPase (1997) J. Gen. Physiol., 109, pp. 555-570
  • 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
  • Bumham, K.P., Anderson, D.R., (2002) Model Selection and Multimodel Inference, 2nd Ed., pp. 60-85. , Springer, New York
  • Smirnova, I.N., Faller, L.D., Mechanism of the conformational change in sodium pump reported by eosin (1995) Biochemistry, 34, pp. 13159-13169
  • Fedosova, N.U., Esmann, M., Nucleotide-binding kinetics of Na, K-ATPase: Cation dependence (2004) Biochemistry, 43, pp. 4212-4218
  • Kaufman, S.B., González-Lebrero, R.M., Rossi, R.C., Garrahan, P.J., Binding of a single Rb+ increases Na+/K +-ATPase activating dephosphorylation without stoichiometric occlusion (2006) J. Biol. Chem., 281, pp. 15721-15726
  • Rossi, R.C., González-Lebrero, R.M., Kaufman, S.B., Garrahan, P.J., Testing K+-like occlusion of cations in the Na +/K+-ATPase (2005) J. Gen. Physiol., 126, pp. 29a

Citas:

---------- APA ----------
Montes, M.R., González-Lebrero, R.M., Garrahan, P.J. & Rossi, R.C. (2006) . Eosin fluorescence changes during Rb+ occlusion in the Na +/K+-ATPase. Biochemistry, 45(43), 13093-13100.
http://dx.doi.org/10.1021/bi060778i
---------- CHICAGO ----------
Montes, M.R., González-Lebrero, R.M., Garrahan, P.J., Rossi, R.C. "Eosin fluorescence changes during Rb+ occlusion in the Na +/K+-ATPase" . Biochemistry 45, no. 43 (2006) : 13093-13100.
http://dx.doi.org/10.1021/bi060778i
---------- MLA ----------
Montes, M.R., González-Lebrero, R.M., Garrahan, P.J., Rossi, R.C. "Eosin fluorescence changes during Rb+ occlusion in the Na +/K+-ATPase" . Biochemistry, vol. 45, no. 43, 2006, pp. 13093-13100.
http://dx.doi.org/10.1021/bi060778i
---------- VANCOUVER ----------
Montes, M.R., González-Lebrero, R.M., Garrahan, P.J., Rossi, R.C. Eosin fluorescence changes during Rb+ occlusion in the Na +/K+-ATPase. Biochemistry. 2006;45(43):13093-13100.
http://dx.doi.org/10.1021/bi060778i