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A K+-selective,three-state channel from fragmented sarcoplasmic reticulum of frog leg muscle
Authors:Pedro P. Labarca  Christopher Miller
Affiliation:(1) Graduate Program in Biophysics and Graduate Department of Biochemistry, Brandeis University, 02254 Waltham, Massachusetts;(2) Present address: Department of Physics, University of California at San Diego, 92093 La Jolla, Calif.
Abstract:Summary Sarcoplasmic reticulum (SR) vesicles from frog leg muscle were fused with a planar phospholipid bilayer by a method described previously for rabbit SR. As a result of the fusion, K+-selective conduction channels are inserted into the bilayer. Unlike the two-state rabbit channel, the frog channel displays three states: a nonconducting (ldquoclosedrdquo) state and two conducting states ldquoagrrdquo and ldquobetardquo. In 0.1m K+ the single-channel conductances are 50 and 150 pS for agr and beta, respectively. The probabilities of appearearance of the three states are voltage-dependent, and transitions between the closed and beta states proceed through the agr state. Both open states follow a quantitatively identical selectivity sequence in channel conductance: K+>NH4+>Rb+>Na+>Li+>Cs+. Both open states are blocked by Cs+ asymmetrically in a voltage-dependent manner. The zero-voltage dissociation constant for blocking is the same for both open states, but the voltage-dependences of the Cs+ block for the two states differ in a way suggesting that the Cs+ blocking site is located more deeply inside the membrane in the beta than in the agr state.
Keywords:Sarcoplasmic reticulum  K-channel, planar bilayer  ion selectivity  Cs-block  excitation-contraction coupling
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