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The principle of gating charge movement in a voltage-dependent K+ channel

Nature volume 423, pages 42–48 (2003)Cite this article

Abstract

The steep dependence of channel opening on membrane voltage allows voltage-dependent K+ channels to turn on almost like a switch. Opening is driven by the movement of gating charges that originate from arginine residues on helical S4 segments of the protein. Each S4 segment forms half of a ‘voltage-sensor paddle’ on the channel's outer perimeter. Here we show that the voltage-sensor paddles are positioned inside the membrane, near the intracellular surface, when the channel is closed, and that the paddles move a large distance across the membrane from inside to outside when the channel opens. KvAP channels were reconstituted into planar lipid membranes and studied using monoclonal Fab fragments, a voltage-sensor toxin, and avidin binding to tethered biotin. Our findings lead us to conclude that the voltage-sensor paddles operate somewhat like hydrophobic cations attached to levers, enabling the membrane electric field to open and close the pore.

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Figure 1: Inhibition of KvAP channels by Fabs and a tarantula venom toxin that bind to the voltage-sensor paddles.
Figure 2: Using avidin and tethered biotin as a molecular ruler to measure positions of the voltage-sensor paddles.
Figure 3: Accessibility of voltage-sensor paddle residues to the internal and external sides of the membrane.
Figure 4: Exposure of the voltage-sensor paddle to the external solution occurs only when the membrane is depolarized.
Figure 5: Positions within the membrane of the voltage-sensor paddles during closed and opened conformations, and a hypothesis for coupling to pore opening.

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Acknowledgements

We thank D. Gadsby and O. Andersen for helpful discussions and advice on the manuscript. This work was supported in part by a grant from the National Institutes of Health (NIH) to R.M. V.R. is supported by a National Science Foundation Graduate Student Research Fellowship, and R.M. is an Investigator in the Howard Hughes Medical Institute.

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Author notes

  1. Youxing Jiang

    Present address: Department of Physiology, University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd, Dallas, Texas, 75390-9040, USA

  2. Roderick MacKinnon: Correspondence and requests for materials should be addressed to R.M.

Authors and Affiliations

  1. Howard Hughes Medical Institute, Laboratory of Molecular Neurobiology and Biophysics, Rockefeller University, 1230 York Avenue, New York, New York, 10021, USA

    Youxing Jiang, Vanessa Ruta, Jiayun Chen, Alice Lee & Roderick MacKinnon

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Correspondence to Roderick MacKinnon.

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Jiang, Y., Ruta, V., Chen, J. et al. The principle of gating charge movement in a voltage-dependent K+ channel. Nature 423, 42–48 (2003). https://doi.org/10.1038/nature01581

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