Voltage-gated ion channels are transmembrane proteins that are essential for nerve impulses and regulate ion flow across cell membranes in response to changes in membrane potential. They are made up of four homologous domains or subunits, each of which contains six transmembrane segments1,2. Studies of potassium channels have shown that the second (S2) and fourth (S4) segments contain several charged residues, which sense changes in voltage and form part of the voltage sensor3,4,5. Although these regions clearly undergo conformational changes in response to voltage6,7,8,9,10, little is known about the nature of these changes because voltage-dependent distance changes have not been measured. Here we use lanthanide-based resonance energy transfer11,12 to measure distances between Shaker potassium channel subunits at specific residues. Voltage-dependent distance changes of up to 3.2 Å were measured at several sites near the S4 segment. These movements directly correlated with electrical measurements of the voltage sensor, establishing the link between physical changes and electrical charge movement. Measured distance changes suggest that the region associated with the S4 segment undergoes a rotation and possible tilt, rather than a large transmembrane movement, in response to voltage. These results demonstrate the first in situ measurement of atomic scale movement in a transmembrane protein.
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This work was supported by NIH grants to F.B. and P.R.S., the Hagiwara Chair funds to F.B., and from Research Corporation to P.R.S. A.C. was also supported by UCLA Medical Scientist Training Program and a National Research Service Award from National Institute of Mental Health. G.E.S. was supported by a National Research Service Award in Molecular Biophysics. We thank A. Gross, for his involvement in preliminary FRET results, and J. Chen, for synthesis of maleimide terbium chelates. We also thank the members of the Bezanilla lab for their support.
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Cha, A., Snyder, G., Selvin, P. et al. Atomic scale movement of the voltage-sensing region in a potassium channel measured via spectroscopy. Nature 402, 809–813 (1999) doi:10.1038/45552
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