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Electron microscopic analysis of KvAP voltage-dependent K+ channels in an open conformation


Voltage-dependent ion channels serve as field-effect transistors by opening a gate in response to membrane voltage changes1. The gate's response to voltage is mediated by voltage sensors2, which are arginine-containing structures that must move with respect to the membrane electric field. We have analysed by electron microscopy a voltage-dependent K+ channel from Aeropyrum pernix (KvAP)3. Fab fragments were attached to ‘voltage sensor paddles’ and identified in the electron microscopy map at 10.5 Å resolution. The extracellular surface location of the Fab fragments in the map is consistent with the membrane-depolarized, open conformation of the channel in electrophysiological experiments. Comparison of the map with a crystal structure4 demonstrates that the voltage sensor paddles are ‘up’ (that is, near the channel's extracellular surface) and situated at the protein–lipid interface. This finding supports the hypothesis that in response to changes in voltage the sensors move at the protein–lipid interface5 rather than in a gating pore surrounded by protein6,7.

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Figure 1: Structure of the KvAP–33H1 complex at 10.5 Å.
Figure 2: Docking of the crystal structures of the KvAP pore and 33H1Fab fragments into the electron microscope map.
Figure 3: Two possible orientations of the paddle relative to the pore.
Figure 4: The S4 probably bends and becomes the S4–S5 linker at Gly 134.

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We thank members of the MacKinnon lab, S. Darst, N. Opalka and D. Stokes for helpful discussions. This work was supported by grants from the NIH to D.N.W. and R.M. R.M is an investigator in the Howard Hughes Medical Institute.

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Correspondence to Roderick MacKinnon.

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Supplementary Figure

Comparison of class averages with projections from the electron microscopy map at representative orientations. (DOC 532 kb)

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Jiang, QX., Wang, DN. & MacKinnon, R. Electron microscopic analysis of KvAP voltage-dependent K+ channels in an open conformation. Nature 430, 806–810 (2004).

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