Abstract
Time-resolved fluorescence and binding studies have been carried out on Na,K-ATPase in the presence of the fluorescent dye eosin Y to obtain thermodynamic and kinetic parameters for the interaction of the enzyme with different cations. Eosin Y binding is indicated by a 3 ns fluorescence decay process and is observed only in the presence of mono- and divalent cations. This type of cation binding is interpreted as a nonselective electrostatic interaction, with negatively charged groups of the enzyme providing a high-affinity eosin Y binding site. Eosin Y binding is observed only under conditions where the enzyme exists in the conformational state F1. The kinetic parameters of eosin Y binding have been determined employing stopped-flow fluorometry.
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References
S. J. D. Karlish (1979) in J. C. Skou and J. G. Nørby (Eds.),Na,K- ATPase Structure and Kinetics, Academic Press, New York, pp. 115–128.
M. L. Helmich-de Jong, J. P. M. van Duynhoven, F. M. A. H. Schuurmans Stekhoven, and J. J. H. H. M. De Pont (1986)Biochim. Biophys. Acta 858, 254–262.
C. Gatto, C. C. Hale, W. Xu, and M. A. Milanick (1995)Biochemistry 34, 965–972.
J. C. Skou and M. Esmann (1981)Biochim. Biophys. Acta 647, 232–240.
J. C. Skou and M. Esmann (1983) inCurrent Topics in Membranes and Transport, Vol. 19, Academic Press, New York, pp. 451–455.
P. L. Jørgensen (1974)Biochim. Biophys. Acta 336, 36–52.
H. Ruf, E. Lewitzki, and E. Grell (1994) in E. Bamberg and W. Schoner (Eds.),The Sodium Pump, Steinkopff, Darmstadt, pp. 569–572.
E. Grell, E. Lewitzki, H. Ruf, and M. Doludda (1994) in E. Bamberg and W. Schoner (Eds.),The Sodium Pump, Steinkopff, Darmstadt, pp. 617–620.
E. Grell, R. Warmuth, E. Lewitzki, H. Ruf (1992)Acta Physiol. Scand. 146, 213–221. ERRATUM:147, 343–344 (1993).
M. Doludda, E. Lewitzki, H. Ruf, and E. Grell (1994) in E. Bamberg and W. Schoner (Eds.),The Sodium Pump, Steinkopff, Darmstadt, pp. 629–632.
E. Lewitzkiet al. unpublished.
M. Esmann (1994)Biochemistry 33, 8558–8565.
E. Grell, E. Lewitzki, and A. Ifftner (1985) in I. M. Glynn and E. Ellory (Eds.),The Sodium Pump, The Company of Biologists Limited, Cambridge, pp. 289–294.
R. M. Fuoss (1958)J. Am. Chem. Soc. 80, 5059–5061.
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Lewitzki, E., Schick, E., Hutterer, R. et al. Conformational changes of Na,K-ATPase probed with eosin Y. J Fluoresc 6, 165–168 (1996). https://doi.org/10.1007/BF00732056
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DOI: https://doi.org/10.1007/BF00732056