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Structure of the calcium pump from sarcoplasmic reticulum at 8-Å resolution

Nature volume 392, pages 835–839 (1998)Cite this article

Abstract

The calcium pump from sarcoplasmic reticulum (Ca2+-ATPase) is typical of the large family of P-type cation pumps. These couple ATP hydrolysis with cation transport, generating cation gradients across membranes. Ca2+-ATPase specifically maintains the low cytoplasmic calcium concentration of resting muscle by pumping calcium into the sarcoplasmic reticulum; subsequent release is used to initiate contraction. No high-resolution structure of a P-type pump has yet been determined, although a 14-Å structure ofCa2+-ATPase, obtained by electron microscopy of frozen-hydrated, tubular crystals1, showed a large cytoplasmic head connected to the transmembrane domain by a narrow stalk. We have now improved the resolution to 8 Å and can discern ten transmembrane α-helices, four of which continue into the stalk. On the basis of constraints from transmembrane topology, site-directed mutagenesis and disulphide crosslinking, we have made tentative assignments for these α-helices within the amino-acid sequence. A distinct cavity leads to the putative calcium-binding site, providing a plausible path for calcium release to the lumen of the sarcoplasmic reticulum.

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Figure 1: Ca2+-ATPase density map at 8-Å resolution.
Figure 2: Density cross-sections through the transmembrane and stalk of Ca2+-ATPase.
Figure 3: Transmembrane architecture of Ca2+-ATPase.
Figure 4: Variability within the transmembrane sequences of P-type ion pumps.
Figure 5: Alternative sequence assignments for the transmembrane helices.

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Acknowledgements

We thank R. Beroukhim and N. Unwin for the use of their programs for helical image analysis, and G. Inesi for the dansyl thapsigargin. This work was partly supported by the NIH and the Ministry of Education, Science, Sports and Culture of Japan.

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Author notes

  1. Koji Yonekura

    Present address: Protonic Nanomachine Project, ERATO, JST, 3-4 Hikaridai, Seika, 619-02, Kyoto, Japan

Authors and Affiliations

  1. Skirball Institute of Biomolecular Medicine, New York University Medical Center, 540 First Avenue, New York, 10016, New York, USA

    Peijun Zhang & David L. Stokes

  2. Institute of Molecular and Cellular Biosciences, The University of Tokyo, Bunkyo-ku, 113, Tokyo, Japan

    Chikashi Toyoshima & Koji Yonekura

  3. National Institute for Medical Research, The Ridgeway, Mill Hill, NW7 1AA, London, UK

    N. Michael Green

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  5. David L. Stokes
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Correspondence to David L. Stokes.

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Zhang, P., Toyoshima, C., Yonekura, K. et al. Structure of the calcium pump from sarcoplasmic reticulum at 8-Å resolution. Nature 392, 835–839 (1998). https://doi.org/10.1038/33959

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