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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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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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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DOI: https://doi.org/10.1038/nsb924
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