Biophys J, August 2001, p. 905-916, Vol. 81, No. 2

Resolving the Gating Charge Movement Associated with Late Transitions in K Channel Activation

Department of Physiology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104 USA

We examined the late transitions in the activation sequence of potassium channels by analyzing gating currents of mutant Shaker IR channels and using the potassium channel blocker 4-aminopyridine (4AP). Gating currents were recorded from a double mutant of Shaker that was nonconducting (W434F mutation) and had the late gating transitions shifted to the right on the voltage axis (L382C mutation), thus separating the late transitions from the early ones. 4AP applied to the double mutant blocked the final transition and made possible novel observations of the isolated intermediate transitions, the ones that immediately precede the final opening of the channel. These transitions, which have not been well characterized previously, produce a distinct fast component in the gating current tails. Two intermediate transitions contribute to the fast component and carry 23% of the total gating charge. The effect of 4AP is well modeled as a selective block of the final gating transition, which opens the channel. The final transition contributes ~5% of the total gating charge.

Biophys J, August 2001, p. 905-916, Vol. 81, No. 2
© 2001 by the Biophysical Society   0006-3495/01/08/905/12  $2.00

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