Letter | Published:

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

Nature volume 409, pages 219223 (11 January 2001) | Download Citation

Subjects

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.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    , & A decade of CLC chloride channels: structure, mechanism, and many unsettled questions. Annu. Rev. Biophys. Biomol. Struct. 29, 411–438 ( 2000).

  2. 2.

    , & High-level expression, functional reconstitution, and quaternary structure of a prokaryotic ClC-type chloride channel. J. Gen. Physiol. 114, 713–722 (1999).

  3. 3.

    & Chloride channels: an emerging molecular picture. BioEssays 19, 117– 126 (1997).

  4. 4.

    , & Two physically distinct pores in the dimeric ClC-0 chloride channel. Nature 383, 340– 343 (1996).

  5. 5.

    , & Independent gating of single pores in CLC-0 chloride channels. Biophys. J. 73, 789– 797 (1997).

  6. 6.

    Open-state substructure of single chloride channels from Torpedo electroplax. Phil. Trans. R. Soc. Lond. B 299, 401– 411 (1982).

  7. 7.

    , & Homodimeric architecture of a ClC-type chloride ion channel. Nature 383, 337– 340 (1996).

  8. 8.

    , & The muscle chloride channel ClC-1 has a double-barreled appearance that is differentially affected in dominant and recessive myotonia. J. Gen. Physiol. 113, 457– 468 (1999).

  9. 9.

    , , & Pore stoichiometry of a voltage-gated chloride channel. Nature 394, 687–690 ( 1998).

  10. 10.

    , & Analysis of electron microscope images and electron diffraction patterns of thin crystals of phi 29 connectors in ice. J. Mol. Biol. 240, 281–287 (1994).

  11. 11.

    , & Purification, reconstitution, and subunit composition of a voltage-gated chloride channel from Torpedo electroplax. Biochemistry 33, 13189–13198 (1994).

  12. 12.

    , & Lipid location in deoxycholate-treated purple membrane at 2.6 Å. J. Mol. Biol. 254, 404–415 (1995).

  13. 13.

    , & Primary structure of Torpedo marmorata chloride channel isolated by expression cloning in Xenopus oocytes. Nature 348, 510–514 ( 1990).

  14. 14.

    , , , & Structure of purple membrane from halobacterium halobium: recording, measurement and evaluation of electron micrographs at 3.5 Å resolution. Ultramicroscopy 19, 147–178 (1986).

  15. 15.

    , & MRC image processing programs. J. Struct. Biol. 116, 9–16 ( 1996).

  16. 16.

    , & Projection structure of rhodopsin. Nature 362, 770–772 ( 1993).

Download references

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.

Author information

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, Massachusetts 02454, USA

    • Nikolaus Grigorieff

Authors

  1. Search for Joseph A. Mindell in:

  2. Search for Merritt Maduke in:

  3. Search for Christopher Miller in:

  4. Search for Nikolaus Grigorieff in:

Corresponding author

Correspondence to Joseph A. Mindell.

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/35051631

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.