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ATP-induced conformational changes of the nucleotide-binding domain of Na,K-ATPase

Nature Structural & Molecular Biology volume 10, pages 468–474 (2003)Cite this article

Abstract

The Na,K-ATPase hydrolyzes ATP to drive the coupled extrusion and uptake of Na+ and K+ ions across the plasma membrane. Here, we report two high-resolution NMR structures of the 213-residue nucleotide-binding domain of rat α1 Na,K-ATPase, determined in the absence and the presence of ATP. The nucleotide binds in the anti conformation and shows a relative paucity of interactions with the protein, reflecting the low-affinity ATP-binding state. Binding of ATP induces substantial conformational changes in the binding pocket and in residues located in the hinge region connecting the N- and P-domains. Structural comparison with the Ca-ATPase stabilized by the inhibitor thapsigargin, E2(TG), and the model of the H-ATPase in the E1 form suggests that the observed changes may trigger the series of events necessary for the release of the K+ ions and/or disengagement of the A-domain, leading to the eventual transfer of the γ-phosphate group to the invariant Asp369.

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Figure 1: Solution structure of the N-domain of NaKα1.
Figure 2: ATP binding to the N-domain of NaKα1.
Figure 3: Docking and superposition of the native (green) and ATP-bound (red) N-domain of NaKα1, the E2(TG) form of Ca-ATPase (violet) and E1 form of H-ATPase (blue).

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Acknowledgements

We thank J.L. van der Plas, J. Plaisier and N. Pannu for setting up software and writing several useful programs, W.J. Rice and co-workers for kindly providing the 11-Å EM map of Na,K-ATPase, R.A.G. de Graaff for critical reading of the manuscript and S. Hilge for professional help with the figures. This work was supported by grants to M.H. of the Swiss National Foundation, the Netherlands Organisation for Scientific Research (NWO) and Technologie stichting STW. G.S. acknowledges the Dutch Royal Academy of Sciences for fellowship support. P.G. acknowledges the Tatsuo Miyazawa Memorial Program of RIKEN for support.

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  1. Mark Hilge

    Present address: Department of Biophysical Chemistry, NSRIM Center, University of Nijmegen, Toernooiveld 1, 6525 ED, Nijmegen, The Netherlands

Authors and Affiliations

  1. Leiden Institute of Chemistry, Gorlaeus Laboratories, Leiden University, P.O. Box 9502, Leiden, 2300, RA, The Netherlands

    Mark Hilge, Gregg Siegal & Jan Pieter Abrahams

  2. Department of Biophysical Chemistry, NSRIM Center, University of Nijmegen, Toernooiveld 1, Nijmegen, 6525, ED, The Netherlands

    Geerten W Vuister

  3. RIKEN Genomic Sciences Center, C112, 1-7-22 Suehiro, Tsurumi, Yokohama, 230-0045, Japan

    Peter Güntert

  4. Institute of Biochemistry, University of Zürich, Winterthurerstr. 190, Zürich, 8057, Switzerland

    Sergio M Gloor

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Correspondence to Mark Hilge.

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Hilge, M., Siegal, G., Vuister, G. et al. ATP-induced conformational changes of the nucleotide-binding domain of Na,K-ATPase. Nat Struct Mol Biol 10, 468–474 (2003). https://doi.org/10.1038/nsb924

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