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Journal of Fluorescence

, Volume 6, Issue 3, pp 165–168 | Cite as

Conformational changes of Na,K-ATPase probed with eosin Y

  • E. Lewitzki
  • E. Schick
  • R. Hutterer
  • F. W. Schneider
  • E. Grell
Article
  • 28 Downloads

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.

Key words

Cation binding fluorescence decay kinetics binding constants Na,K-ATPase eosin Y 

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Copyright information

© Plenum Publishing Corporation 1996

Authors and Affiliations

  • E. Lewitzki
    • 1
  • E. Schick
    • 1
  • R. Hutterer
    • 2
  • F. W. Schneider
    • 2
  • E. Grell
    • 1
  1. 1.Max-Planck-Institute of BiophysicsFrankfurtGermany
  2. 2.Institute of Physical ChemistryUniversity of WürzburgWürzburgGermany

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