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Gating and Conductance Changes in BK Ca Channels in Bilayers Are Reciprocal

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Abstract

The energy associated with a mismatch between the hydrocarbon portions of a lipid bilayer and the hydrophobic regions of a transmembrane protein requires that one or both components deform in an attempt to minimize the energy difference. Transmembrane potassium channel subunits are composed of different structural motifs, each responsible for ion-selectivity, conductance and gating capabilities. Each has an inherent degree of flexibility commensurate with its amino acid composition. It is not clear, however, how each structural motif will respond to a fixed amount of distortion applied to the whole structure. We examined the single-channel conductance (Gc) and gating (open probability, P o) of single BKCa channels (hslo α-subunits) inserted into planar lipid bilayers containing 1,2-dioleoyl-3-phosphatidylethanolamine (DOPE) or DOPE with either 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) or sphingomyelin (SPM) and 1-palmitoyl-2-oleoyl-3-phosphatidylethanolamine (POPE) with SPM. These latter three binary mixtures formed stable membranes with different distributions of thickness domains as determined by atomic force microscopy. Channels placed in each composition should be exposed to different amounts of distortion. BKCa channels forced into the DOPE/SPM bilayer containing lipid domains with two different thicknesses showed two distinct levels of Gc and Po. The alterations in Gc and Po were reciprocal. A larger conductance was accompanied by a smaller value for gating and vice versa. Channels forced into the POPE/SPM bilayer containing lipid domains with different thicknesses showed more than two distinct levels of Gc and Po. Channels placed in a uniform bilayer (DOPE/DOPC) showed a uniform distribution of conductance and activation. We conclude that both the inner and outer domains of the channel where these two channel functions are localized respond to deformation and that a fixed amount of distortion results in reciprocal changes in protein function.

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Acknowledgement

We thank Paula Zadek and Andrew Wilson for assistance with growing the HEK-293 cells and John Crowley for helpful discussions and assistance with the preparation of BKCa channels containing membrane fragments from them. Funds were provided by National Institutes of Health grant AA12054 (to S. N. T.).

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  1. Brudnick Neuropsychiatric Research Institute, University of Massachusetts Medical School, 303 Belmont Street, Worcester, Massachusetts, 01604

    Robert J. O’Connell, Chunbo Yuan & Steven N. Treistman

  2. Steacie Institute for Molecular Sciences, National Research Council of Canada, 100 Sussex Drive, Ottawa, Ontario, K1A 0R6, Canada

    Linda J. Johnston, Olga Rinco & Ira Probodh

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  1. Robert J. O’Connell
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  2. Chunbo Yuan
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O’Connell, R.J., Yuan, C., Johnston, L.J. et al. Gating and Conductance Changes in BK Ca Channels in Bilayers Are Reciprocal. J Membrane Biol 213, 143–153 (2006). https://doi.org/10.1007/s00232-006-0034-1

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