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Single anion channels reconstituted from cardiac mitoplasts

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Abstract

Ion channels from sheep cardiac mitoplast (inverted inner mitochondrial membrane vesicle) preparations were incorporated into voltage-clamped planar lipid bilayers. The appearance of anion rather than cation channels could be promoted by exposing the bilayers to osmotic gradients formed by Cl− salts of large, relatively imperment, cations at a pH of 8.8. Two distinct activities were identified. These comprised a multisubstate anion channel of intermediate conductance (∼60 pS in 300vs. 50mm choline Cl, ∼100 pS in symmetric 150mm KCl), and a lower-conductance anion channel (∼25 or ∼50 pS in similar conditions), which only displayed two well-defined substates, at ∼25 and ∼50% of the fully open state. The larger channels were not simple multiples of the lower-conductance channels, but both discriminated poorly, and to a similar extent, between anions and cations (PCl −/Pcholine + ∼12, PCl −/PK +∼8). The lower-conductance channel was only minimally selective between different anions (PNO −3 (1.0)=PCl −>PBr −>PI −>PSCN −(0.8)), and its conductance failed to saturate even in high (>1.0 M) activities of KCl. The channels were not obviously voltage dependent, and they were unaffected by 0.5 mM SITS, H2O2, propranolol, quinine or amitriptyline, or by 2 mM ATP, or by variations in pH (5.5–8.8). Ca2+ and Mg2+ did not alter single channel activity, but did modify single current amplitudes in the lower-conductance channel. This effect, together with voltage-dependent substate behavior, is described in the following paper.

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  1. Department of Biochemistry, University of Edinburgh, George Square, EH8 9XD, Edinburgh, Scotland, UK

    K. A. Hayman, T. D. Spurway & R. H. Ashley

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  1. K. A. Hayman
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  2. T. D. Spurway
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  3. R. H. Ashley
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Hayman, K.A., Spurway, T.D. & Ashley, R.H. Single anion channels reconstituted from cardiac mitoplasts. J. Membrain Biol. 136, 181–190 (1993). https://doi.org/10.1007/BF02505762

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