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Surface Charges of the Membrane Crucially Affect Regulation of Na,K-ATPase by Phospholemman (FXYD1)

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Abstract

The human α1/His10-β1 isoform of Na,K-ATPase has been reconstituted as a complex with and without FXYD1 into proteoliposomes of various lipid compositions in order to study the effect of the regulatory subunit on the half-saturating Na+ concentration (K 1/2) of Na+ ions for activation of the ion pump. It has been shown that the fraction of negatively charged lipid in the bilayer crucially affects the regulatory properties. At low concentrations of the negatively charged lipid DOPS (<10 %), FXYD1 increases K 1/2 of Na+ ions for activation of the ion pump. Phosphorylation of FXYD1 by protein kinase A at Ser68 abrogates this effect. Conversely, for proteoliposomes made with high concentrations of DOPS (>10 %), little or no effect of FXYD1 on the K 1/2 of Na+ ions is observed. Depending on ionic strength and lipid composition of the proteoliposomes, FXYD1 can alter the K 1/2 of Na+ ions by up to twofold. We propose possible molecular mechanisms to explain the regulatory effects of FXYD1 and the influence of charged lipid and protein phosphorylation. In particular, the positively charged C-terminal helix of FXYD1 appears to be highly mobile and may interact with the cytoplasmic N domain of the α-subunit, the interaction being strongly affected by phosphorylation at Ser68 and the surface charge of the membrane.

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Acknowledgments

We thank Dr. Kay Diederichs for his help in creating the figures with the molecular structures. The work was financially supported by the German Israeli Foundation (Grant 922-165.9 to H.-J. A. and S. J. K.)

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  1. Erica Cirri

    Present address: Molecular Mechanisms of Membrane Transport Laboratory, Institut Pasteur, 25 rue du Docteur Roux, 75724, Paris Cedex 15, France

Authors and Affiliations

  1. Department of Biology and Graduate School of Chemical Biology, University of Konstanz, 78464, Constance, Germany

    Erica Cirri, Corinna Kirchner, Simon Becker & Hans-Jürgen Apell

  2. Department of Biological Chemistry, Weizmann Institute of Science, 76100, Rehovot, Israel

    Adriana Katz & Steven J. Karlish

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  1. Erica Cirri
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  2. Corinna Kirchner
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Correspondence to Hans-Jürgen Apell.

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Cirri, E., Kirchner, C., Becker, S. et al. Surface Charges of the Membrane Crucially Affect Regulation of Na,K-ATPase by Phospholemman (FXYD1). J Membrane Biol 246, 967–979 (2013). https://doi.org/10.1007/s00232-013-9600-5

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