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Crystal structure of a voltage-gated sodium channel in two potentially inactivated states

Nature volume 486, pages 135–139 (2012)Cite this article

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Abstract

In excitable cells, voltage-gated sodium (NaV) channels activate to initiate action potentials and then undergo fast and slow inactivation processes that terminate their ionic conductance1,2. Inactivation is a hallmark of NaV channel function and is critical for control of membrane excitability3, but the structural basis for this process has remained elusive. Here we report crystallographic snapshots of the wild-type NaVAb channel from Arcobacter butzleri captured in two potentially inactivated states at 3.2 Å resolution. Compared to previous structures of NaVAb channels with cysteine mutations in the pore-lining S6 helices (ref. 4), the S6 helices and the intracellular activation gate have undergone significant rearrangements: one pair of S6 helices has collapsed towards the central pore axis and the other S6 pair has moved outward to produce a striking dimer-of-dimers configuration. An increase in global structural asymmetry is observed throughout our wild-type NaVAb models, reshaping the ion selectivity filter at the extracellular end of the pore, the central cavity and its residues that are analogous to the mammalian drug receptor site, and the lateral pore fenestrations. The voltage-sensing domains have also shifted around the perimeter of the pore module in wild-type NaVAb, compared to the mutant channel, and local structural changes identify a conserved interaction network that connects distant molecular determinants involved in NaV channel gating and inactivation. These potential inactivated-state structures provide new insights into NaV channel gating and novel avenues to drug development and therapy for a range of debilitating NaV channelopathies.

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Figure 1: Structure and function of Na V Ab.
Figure 2: Structural changes in the selectivity filter of Na V Ab.
Figure 3: Conformational changes in the pore module of Na V Ab.
Figure 4: Structure and coupling of the VSD in Na V Ab.

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Protein Data Bank

Data deposits

Coordinates and structure factors have been deposited in the Protein Data Bank under accession code 4EKW.

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Acknowledgements

We thank B. Hille for comments on a draft of the manuscript and members of the Zheng and Catterall groups for their insight and support throughout this project. We are grateful to the beamline staff at the Advanced Light Source (BL8.2.1 and BL8.2.2) for their assistance during data collection. J.P. acknowledges support from a Canadian Institutes of Health Research fellowship and the support of N. and E. Payandeh. This work was supported by grants from the National Institutes of Health (R01 NS15751 and U01 NS058039 to W.A.C.) and by the Howard Hughes Medical Institute (N.Z.). We dedicate this work to the memory of our colleague Laura Sheard.

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Author notes

  1. Jian Payandeh

    Present address: Present address: Department of Structural Biology, Genentech, Inc., South San Francisco, California 94080, USA.,

Authors and Affiliations

  1. Department of Pharmacology, University of Washington, Seattle, 98195, Washington, USA

    Jian Payandeh, Tamer M. Gamal El-Din, Todd Scheuer, Ning Zheng & William A. Catterall

  2. Howard Hughes Medical Institute, University of Washington, Seattle, Washington, 98195, USA

    Ning Zheng

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Contributions

W.A.C. and N.Z. are both senior authors. J.P., N.Z. and W.A.C. conceived and J.P. conducted the protein purification and crystallization experiments. J.P. and N.Z. determined and analysed the structure of NaVAb. T.M.G., T.S. and W.A.C. conceived, T.M.G. conducted, and T.M.G., T.S. and W.A.C. analysed the electrophysiology experiments. All authors contributed to writing the manuscript.

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Correspondence to Ning Zheng or William A. Catterall.

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The authors declare no competing financial interests.

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Payandeh, J., Gamal El-Din, T., Scheuer, T. et al. Crystal structure of a voltage-gated sodium channel in two potentially inactivated states. Nature 486, 135–139 (2012). https://doi.org/10.1038/nature11077

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