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The structural basis of calcium transport by the calcium pump

Nature volume 450, pages 1036–1042 (2007)Cite this article

Abstract

The sarcoplasmic reticulum Ca2+-ATPase, a P-type ATPase, has a critical role in muscle function and metabolism. Here we present functional studies and three new crystal structures of the rabbit skeletal muscle Ca2+-ATPase, representing the phosphoenzyme intermediates associated with Ca2+ binding, Ca2+ translocation and dephosphorylation, that are based on complexes with a functional ATP analogue, beryllium fluoride and aluminium fluoride, respectively. The structures complete the cycle of nucleotide binding and cation transport of Ca2+-ATPase. Phosphorylation of the enzyme triggers the onset of a conformational change that leads to the opening of a luminal exit pathway defined by the transmembrane segments M1 through M6, which represent the canonical membrane domain of P-type pumps. Ca2+ release is promoted by translocation of the M4 helix, exposing Glu 309, Glu 771 and Asn 796 to the lumen. The mechanism explains how P-type ATPases are able to form the steep electrochemical gradients required for key functions in eukaryotic cells.

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Figure 1: Overall comparison of SERCA1a structures representing key states of the reaction cycle.
Figure 2: The Ca2E1∼P state obtained with AMPPNP.
Figure 3: The E2P state obtained with beryllium fluoride.
Figure 4: The luminal exit pathway of sarcoplasmic reticulum Ca 2+ -ATPase.
Figure 5: Changes at the phosphorylation site of SERCA1a in the functional cycle.
Figure 6: Schematic representation of the reaction cycle.

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Acknowledgements

We dedicate this paper to the memory of B. Holm. We thank B. Nielsen, M.-B. Hemmingsen and A. M. Nielsen for technical assistance; J. L. Karlsen and F. Fredslund for technical discussions; and D. Flot and L. Gordon at beamlines ID 23-1 and -2 (operated jointly with EMBL-Grenoble) and ID 29 at the European Synchrotron Radiation Facility (ESRF) for help with data collection. Beamtime at the EMBL-DESY synchrotron Hamburg Germany is also acknowledged. This work was supported by the Danish Natural Science Research Council through the DANSYNC program, the Danish Medical Research Council, the Aarhus University Research Foundation, and the Novo Nordisk Foundation. C.Ol. is the recipient of a stipend from the PC Petersen Foundation and a PhD fellowship from the faculty of Health Sciences Aarhus University. A PhD fellowship (A.-M.L.W.) was financed by the Lundbeck Foundation. M.P. was supported by a post-doctoral fellowship from the Federation of European Biochemical Societies (FEBS) and P.N. is supported by a Hallas-Møller stipend of the Novo Nordisk Foundation.

Author Contributions C.Ol., M.P., A.-M.L.W., J.V.M. and P.N. contributed equally to this work. J.P.M. assisted with data collection and structure determination. C.Ox. and C.G. contributed with mass-spectrometry data and analysis.

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  1. Martin Picard

    Present address: Present address: Laboratoire de Cristallographie et RMN biologiques, UMR 8015 CNRS, Faculté de Pharmacie. Université Paris Descartes, 4 avenue de l'Observatoire, 75270 Paris Cedex 06, France.,

Authors and Affiliations

  1. Centre for Membrane Pumps in Cells and Disease—PUMPKIN, Danish National Research Foundation, and,,

    Claus Olesen, Anne-Marie Lund Winther, J. Preben Morth, Jesper Vuust Møller & Poul Nissen

  2. Institute of Physiology and Biophysics, University of Aarhus, Ole Worms Alle, blg. 1185, DK - 8000 Aarhus C, Denmark ,

    Claus Olesen & Jesper Vuust Møller

  3. Department of Molecular Biology, University of Aarhus, Gustav Wieds Vej 10C, DK - 8000 Aarhus C, Denmark,

    Martin Picard, Anne-Marie Lund Winther, Claus Gyrup, J. Preben Morth, Claus Oxvig & Poul Nissen

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Additional information

The structural data have been deposited with the following codes in the Protein Data Bank: Ca2E1∼P, 3BA6; E2-AlF4-, 3B9R; and E2-BeF3-, 3B9B.

Supplementary information

Supplementary Information

The file contains Supplementary Table 1, Supplementary Figures S1-S6 with Legends and additional references. (PDF 1784 kb)

Supplementary Movie

The file contains Supplementary Movie 1. The movie shows the structure of the α β γ-complex of the Na,K-ATPase from pig kidney rotating (α-subunit in blue, β in wheat, γ in red, C-terminal switch in pink, and two Rb+ ions in magenta. The β ectodomain is represented by the experimental electron density (MOV 3297 kb)

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Olesen, C., Picard, M., Winther, AM. et al. The structural basis of calcium transport by the calcium pump. Nature 450, 1036–1042 (2007). https://doi.org/10.1038/nature06418

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