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Projection structure of a ClC-type chloride channel at 6.5 Å resolution

Nature volume 409, pages 219–223 (2001)Cite this article

Abstract

Virtually all cells in all eukaryotic organisms express ion channels of the ClC type, the only known molecular family of chloride-ion-selective channels. The diversity of ClC channels highlights the multitude and range of functions served by gated chloride-ion conduction in biological membranes, such as controlling electrical excitability in skeletal muscle, maintaining systemic blood pressure, acidifying endosomal compartments, and regulating electrical responses of GABA (γ-aminobutyric acid)-containing interneurons in the central nervous system1. Previously, we expressed and purified a prokaryotic ClC channel homologue2. Here we report the formation of two-dimensional crystals of this ClC channel protein reconstituted into phospholipid bilayer membranes. Cryo-electron microscopic analysis of these crystals yields a projection structure at 6.5 Å resolution, which shows off-axis water-filled pores within the dimeric channel complex.

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Figure 1: Active 2D crystals of EriC.
Figure 2: Analysis of glucose-embedded EriC crystals.
Figure 3: Quality of diffraction data.
Figure 4: Projection density map of EriC at 6.5 Å resolution.

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Acknowledgements

We are grateful to R. Henderson for making facilities at the MRC, including the Hitachi microscope, available to us for low-dose experiments, to J. Berriman for help and hand-holding, and to T. Walz for crucial advice on crystallization. We also acknowledge D. Pheasant and L. Melanson for technical assistance. Support was provided in part by a grant to J.A.M. from the NIH.

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Authors and Affiliations

  1. Department of Biochemistry, Howard Hughes Medical Institute,

    Joseph A. Mindell, Merritt Maduke, Christopher Miller & Nikolaus Grigorieff

  2. Rosenstiel Basic Medical Science Research Center and W. M. Keck Institute for Cellular Visualization, Brandeis University, Waltham, 02454, Massachusetts, USA

    Nikolaus Grigorieff

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  1. Joseph A. Mindell
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  2. Merritt Maduke
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Correspondence to Joseph A. Mindell.

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Mindell, J., Maduke, M., Miller, C. et al. Projection structure of a ClC-type chloride channel at 6.5 Å resolution. Nature 409, 219–223 (2001). https://doi.org/10.1038/35051631

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