title: Voltage dependence of the Ca(2+)-activated K(+) channel K(Ca)3.1 in human erythroleukemia cells.
abstract: We have isolated a K(+)-selective, Ca(2+)-dependent whole cell current and single-channel correlate in the human erythroleukemia (HEL) cell line. The whole cell current was inhibited by the intermediate-conductance KCa3.1 inhibitors clotrimazole, TRAM-34, and charybdotoxin, unaffected by the small-conductance KCa2 family inhibitor apamin and the large-conductance KCa1.1 inhibitors paxilline and iberiotoxin, and augmented by NS309. The single-channel correlate of the whole cell current was blocked by TRAM-34 and clotrimazole, insensitive to paxilline, and augmented by NS309 and had a single-channel conductance in physiological K(+) gradients of ~9 pS. RT-PCR revealed that the KCa3.1 gene, but not the KCa1.1 gene, was expressed in HEL cells. The KCa3.1 current, isolated in HEL cells under whole cell patch-clamp conditions, displayed an activated current component during depolarizing voltage steps from hyperpolarized holding potentials and tail currents upon repolarization, consistent with voltage-dependent modulation. This activated current increased with increasing voltage steps above -40 mV and was sensitive to inhibition by clotrimazole, TRAM-34, and charybdotoxin and insensitive to apamin, paxilline, and iberiotoxin. In single-channel experiments, depolarization resulted in an increase in open channel probability (Po) of KCa3.1, with no increase in channel number. The voltage modulation of Po was an increasing monotonic function of voltage. In the absence of elevated Ca(2+), voltage was ineffective at inducing channel activity in whole cell and single-channel experiments. These data indicate that KCa3.1 in HEL cells displays a unique form of voltage dependence modulating Po.
authors: Department of Physiology, Development, and Neuroscience, University of Cambridge, Cambridge, United Kingdom.
authors: Colin J
authors: CJ
authors: Stoneking
authors: Oshini
authors: O
authors: Shivakumar
authors: David Nicholson
authors: DN
authors: Thomas
authors: William H
authors: WH
authors: Colledge
authors: Michael J
authors: MJ
authors: Mason
mesh_terms: D002118:Calcium / Q000502:physiology; D045744:Cell Line, Tumor; D018999:Charybdotoxin / Q000494:pharmacology; D003022:Clotrimazole / Q000494:pharmacology; D004558:Electric Stimulation; D015870:Gene Expression; D006801:Humans; D051660:Intermediate-Conductance Calcium-Activated Potassium Channels / Q000235:genetics / Q000378:metabolism*; D015640:Ion Channel Gating; D051037:Large-Conductance Calcium-Activated Potassium Channel alpha Subunits / Q000235:genetics / Q000378:metabolism; D004915:Leukemia, Erythroblastic, Acute; D008274:Magnesium / Q000502:physiology; D008564:Membrane Potentials / Q000187:drug effects; D018408:Patch-Clamp Techniques; D011188:Potassium / Q000502:physiology; D026902:Potassium Channel Blockers / Q000494:pharmacology; D011720:Pyrazoles / Q000494:pharmacology
chemical_list: D051660:Intermediate-Conductance Calcium-Activated Potassium Channels; C494841:KCNMA1 protein, human; C496923:KCNN4 protein, human; D051037:Large-Conductance Calcium-Activated Potassium Channel alpha Subunits; D026902:Potassium Channel Blockers; D011720:Pyrazoles; C411671:TRAM 34; D018999:Charybdotoxin; D003022:Clotrimazole; D008274:Magnesium; D011188:Potassium; D002118:Calcium
keywords: KCa3.1; open channel probability; potassium channel; voltage dependence