A Ba2+ chelator suppresses long shut events in fully activated high-conductance Ca(2+)-dependent K+ channels.
High-conductance Ca(2+)-activated K+ channels from rat skeletal muscle were incorporated into planar lipid bilayers, and the channel kinetics were studied with a high internal Ca2+ concentration (Cai). Raising the Cai is known to increase the channel open probability. This effect is due to an increases in openings frequency and duration, and saturates at a Cai around 100 microM. Raising the Cai also increases the occurrence of less frequent but very long (> 5 s) shut events. The mechanism underlying this slow kinetic process was studied. Raising Cai above 100 microM does not further increase the frequency of the long shut events. This was not consistent with the hypothesis that the long closures result from a classical channel-block mechanism induced by internal Ca2+. The transmembrane voltage and the presence of K+ ions in the external compartment both affect the slow kinetic process. A comparison of these effects with the properties of the channel block induced by Ba2+ ions added to the internal compartment strongly suggested that the long shut events are due to a contamination of the internal solutions by Ba2+. This was confirmed by showing that a crown-ether compound that strongly chelates Ba2+ completely suppresses the long shut events when added to the inner compartment.
Neyton
J
J
Laboratoire de Neurobiologie, Ecole Normale Supérieure, CNRS URA 1857, Paris, France. neyton@wotan.ens.fr
D000818:Animals; D001464:Barium / Q000494:pharmacology*; D055592:Biophysical Phenomena; D001703:Biophysics; D002118:Calcium / Q000378:metabolism*; D002614:Chelating Agents / Q000494:pharmacology*; D043844:Crown Ethers; D004553:Electric Conductivity; D004988:Ethers, Cyclic / Q000494:pharmacology; D066298:In Vitro Techniques; D007700:Kinetics; D051036:Large-Conductance Calcium-Activated Potassium Channels; D008051:Lipid Bilayers / Q000378:metabolism; D018482:Muscle, Skeletal / Q000378:metabolism; D015221:Potassium Channels / Q000187:drug effects* / Q000378:metabolism*; D024681:Potassium Channels, Calcium-Activated; D051381:Rats
D002614:Chelating Agents; D043844:Crown Ethers; D004988:Ethers, Cyclic; D051036:Large-Conductance Calcium-Activated Potassium Channels; D008051:Lipid Bilayers; D015221:Potassium Channels; D024681:Potassium Channels, Calcium-Activated; C090732:18-crown-6 2,3,11,12-tetracarboxylic acid; D001464:Barium; D002118:Calcium