ArticleBiophysical journal1994
Reconstitution of expressed KCa channels from Xenopus oocytes to lipid bilayers.
Article in Biophysical journal, 1994. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed, 55 citations in OpenAlex.
- dyschronic, a Drosophila homolog of a deaf-blindness gene, regulates circadian output and Slowpoke channels.PLoS genetics · 2012Article
- Splicing of the rSlo gene affects the molecular composition and drug response of Ca2+-activated K+ channels in skeletal muscle.PloS one · 2012Article
- In vivo phosphorylation of CFTR promotes formation of a nucleotide-binding domain heterodimer.The EMBO journal · 2006Article
- Bilayer reconstitution of voltage-dependent ion channels using a microfabricated silicon chip.Biophysical journal · 2001Article
- A novel type of internal barium block of a maxi-K+ channel from human vas deferens epithelial cells.Biophysical journal · 1998Article
- A novel calcium-sensing domain in the BK channel.Biophysical journal · 1997Article
- Allosteric gating of a large conductance Ca-activated K+ channel.The Journal of general physiology · 1997Article
- Molecular constituents of maxi KCa channels in human coronary smooth muscle: predominant alpha + beta subunit complexes.The Journal of physiology · 1997Article
- Intrinsic voltage dependence and Ca2+ regulation of mslo large conductance Ca-activated K+ channels.The Journal of general physiology · 1997Article
- Point mutations in alpha bENaC regulate channel gating, ion selectivity, and sensitivity to amiloride.Biophysical journal · 1997Article
- Cloning, expression, and distribution of a Ca(2+)-activated K+ channel beta-subunit from human brain.Proceedings of the National Academy of Sciences of the United States of America · 1996Article
- Interaction of internal Ba2+ with a cloned Ca(2+)-dependent K+ (hslo) channel from smooth muscle.The Journal of general physiology · 1996Article
- Two barium binding sites on a maxi K+ channel from human vas deferens epithelial cells.Biophysical journal · 1996Article
- Potentiation of large conductance KCa channels by niflumic, flufenamic, and mefenamic acids.Biophysical journal · 1994Article
Corrections and comments
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
Abstract
Reconstitution of large conductance calcium-activated potassium (KCa) channels from native cell membranes into planar lipid bilayers provides a powerful method to study single channel properties, including ion conduction, pharmacology, and gating. Recently, KCa channels derived from the Drosophila Slowpoke (Slo) gene have been cloned and heterologously expressed in Xenopus oocytes. In this report, we describe the reconstitution of cloned and expressed Slo KCa channels from Xenopus oocyte membranes into lipid bilayers. The reconstituted channels demonstrate functional properties characteristic of native KCa channels. They possess a mean unitary conductance of approximately 260 pS in symmetrical potassium (250 mM), and they are voltage- and calcium-sensitive. At 50 microM Ca2+, their half-activation potential was near -20 mV; and their affinity for calcium is in the micromolar range. Reconstituted Slo KCa channels were insensitive to external charybdotoxin (40-500 nM) and sensitive to micromolar concentrations of external tetraethylammonium (KD = 158 microM, at 0 mV) and internal Ba2+ (KD = 76 microM, at 40 mV). In addition, they were blocked by internally applied "ball" inactivating peptide (KD = 480 microM, at 40 mV). These results demonstrate that cloned KCa channels expressed in Xenopus oocytes can be readily incorporated into lipid bilayers where detailed mechanistic studies can be performed under controlled internal and external experimental conditions.
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