Ion Selectivity of the cytoplasmic binding sites of the Na,K-ATPase : I. sodium binding is associated with a conformational rearrangement

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1999
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Schneeberger, Anne
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Journal of Membrane Biology. 1999, 168(3), pp. 221-228. ISSN 0022-2631. eISSN 1432-1424. Available under: doi: 10.1007/s002329900511
Zusammenfassung

To investigate Na+ binding to the ion-binding sites presented on the cytoplasmic side of the Na,KATPase, equilibrium Na+-titration experiments were performed using two fluorescent dyes, RH421¹ and FITC, to detect protein-specific actions. Fluorescence changes upon addition of Na+ in the presence of various Mg2+ concentrations were similar and could be fitted with a Hill function. The half-saturating concentrations and Hill coefficients determined were almost identical. As RH421 responds to binding of a Na+ ion to the third neutral site whereas FITC monitors conformational changes in the ATP-binding site or its environment, this result implies that electrogenic binding of the third Na+ ion is the trigger for a structural rearrangement of the ATP-binding moiety. This enables enzyme phosphorylation, which is accompanied by a fast occlusion of the Na+ ions and followed by the conformational transition E1/E2 of the protein. The coordinated action both at the ion and the nucleotide binding sites allows for the first time a detailed formulation of the mechanism of enzyme phosphorylation that occurs only when three Na+ ions are bound.

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Fachgebiet (DDC)
570 Biowissenschaften, Biologie
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Na,K-ATPase, Cytoplasmic ion binding, Electrochromic fluorescent dye, FITC, Ion transport, Energy transduction mechanism
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ISO 690SCHNEEBERGER, Anne, Hans-Jürgen APELL, 1999. Ion Selectivity of the cytoplasmic binding sites of the Na,K-ATPase : I. sodium binding is associated with a conformational rearrangement. In: Journal of Membrane Biology. 1999, 168(3), pp. 221-228. ISSN 0022-2631. eISSN 1432-1424. Available under: doi: 10.1007/s002329900511
BibTex
@article{Schneeberger1999Selec-7625,
  year={1999},
  doi={10.1007/s002329900511},
  title={Ion Selectivity of the cytoplasmic binding sites of the Na,K-ATPase : I. sodium binding is associated with a conformational rearrangement},
  number={3},
  volume={168},
  issn={0022-2631},
  journal={Journal of Membrane Biology},
  pages={221--228},
  author={Schneeberger, Anne and Apell, Hans-Jürgen}
}
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