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Molecular determinants of coupling between the domain III voltage sensor and pore of a sodium channel

Nature Structural & Molecular Biology volume 17, pages 230–237 (2010)Cite this article

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Abstract

In a voltage-dependent sodium channel, the activation of voltage sensors upon depolarization leads to the opening of the pore gates. To elucidate the principles underlying this conformational coupling, we investigated a putative gating interface in domain III of the sodium channel using voltage-clamp fluorimetry and tryptophan-scanning mutagenesis. Most mutations have similar energetic effects on voltage-sensor activation and pore opening. However, several mutations stabilized the activated voltage sensor while concurrently destabilizing the open pore. When mapped onto a homology model of the sodium channel, most localized to hinge regions of the gating interface. Our analysis shows that these residues are involved in energetic coupling of the voltage sensor to the pore when both are in resting and when both are in activated conformations, supporting the notion that electromechanical coupling in a voltage-dependent ion channel involves the movement of rigid segments connected by elastic hinges.

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Figure 1: Membrane topology of a sodium channel and a sequence comparison of the skeletal muscle sodium channel with the Shaker and Kv1.2/2.1 chimeric potassium channel.
Figure 2: Voltage-dependent conductance and fluorescence responses in the mutant sodium channels.
Figure 3: Superposition of the homology model of sodium channel domain III with the structure of the Kv1.2/2.1 chimera and mapping the prominent class I positions on the homology model.
Figure 4: Normalized G-V (blue) and F-V relationships (red) for prominent class II mutants.
Figure 5: Mapping the prominent class II mutants on to the structural model of sodium channel DIII.
Figure 6: The G-V and F-V curves upon perturbation of putative interaction pairs and mapping of the positions of an interaction pair onto the homology model of sodium channel DIII.
Figure 7: Two general models describing the coupled activation process involving the voltage sensor and pore of a voltage-dependent ion channel.

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Acknowledgements

This project was supported by funds from the US National Institutes of Health (GM084140-01), the American Heart Association Scientist Development Award (0535214N) and the Shaw Scientist award to B.C. We thank M. Goldschen for help with preparing Supplementary Figure 3, K. Schuldt for excellent technical assistance and the members of the Chanda laboratory for their comments and discussions.

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Author notes

  1. Sandipan Chowdhury

    Present address: Biophysics Graduate Training Program, University of Wisconsin, Madison, Wisconsin, USA

  2. Yukiko Muroi

    Present address: Present address: Division of Allergy and Clinical Immunology, Department of Medicine, Johns Hopkins University, Baltimore, Maryland, USA.,

  3. Yukiko Muroi and Manoel Arcisio-Miranda: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Physiology, School of Medicine and Public Health, University of Wisconsin, Madison, Wisconsin, USA

    Yukiko Muroi, Manoel Arcisio-Miranda, Sandipan Chowdhury & Baron Chanda

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Contributions

Y.M. and M.A-.M. contributed equally to this work; Y.M. and M.A.-M. performed all the experiments and analyzed the data; S.C. created the structural model; S.C. and B.C. generated the coupling models; Y.M., M.A.-M. and B.C. wrote the paper.

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Correspondence to Baron Chanda.

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Supplementary Figures 1–4, Supplementary Tables 1 and 2, and Supplementary Methods (PDF 2066 kb)

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Muroi, Y., Arcisio-Miranda, M., Chowdhury, S. et al. Molecular determinants of coupling between the domain III voltage sensor and pore of a sodium channel. Nat Struct Mol Biol 17, 230–237 (2010). https://doi.org/10.1038/nsmb.1749

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