1. Characterisation of large-conductance calcium-activated potassium channels (BK(Ca)) in human NT2-N cells
- Author
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Peter W. Andrews, Hugh Chapman, C. Piggot, and Kenneth Taylor Wann
- Subjects
Time Factors ,Potassium ,chemistry.chemical_element ,Membrane Potentials ,chemistry.chemical_compound ,symbols.namesake ,Adenosine Triphosphate ,Cell Line, Tumor ,Potassium Channel Blockers ,Reaction Time ,Humans ,Patch clamp ,Calcium Signaling ,Large-Conductance Calcium-Activated Potassium Channels ,Paxilline ,Molecular Biology ,Neurons ,Hill differential equation ,Dose-Response Relationship, Drug ,General Neuroscience ,Cell Membrane ,Time constant ,Conductance ,Calcium-activated potassium channel ,Potassium channel ,chemistry ,Biochemistry ,Biophysics ,symbols ,Calcium ,Neurology (clinical) ,Ion Channel Gating ,Developmental Biology - Abstract
Large-conductance calcium-activated potassium (BK(Ca)) channels were studied in inside-out patches of human NTERA2 neuronal cells (NT2-N). In symmetrical (140 mM) K(+) the channel mean conductance was 265 pS, the current reversing at approximately 0 mV. It was selective (P(K)/P(Na)=20:1) and blocked by internal paxilline and TEA. The open probability-voltage relationship for BK(Ca) was fitted with a Boltzmann function, the V((1/2)) being 76.3 mV, 33.6 mV and -14.1 mV at 0.1 muM, 3.3 muM and 10 muM [Ca(2+)](i), respectively. The relationship between open probability and [Ca(2+)](i) was fitted by the Hill equation (Hill coefficient 2.7, half maximal activation at 2.0 muM [Ca(2+)](i)). Open and closed dwell time histograms were fitted by the sum of two and three voltage-dependent exponentials, respectively. Increasing [Ca(2+)](i) produced both an increase in the longer open time constant and a decrease in the longest closed time constant, so increasing mean open time. "Intracellular" ATP evoked a concentration-dependent increase in NT2-N BK(Ca) activity. At +40 mV half-maximum activation occurred at an [ATP](i) of 3 mM (30 nM [Ca(2+)](i)). ADP and GTP were less potent, and AMP-PNP was inactive. This is the first characterisation of a potassium channel in NT2-N cells showing that it is similar to the BK(Ca) channel of other preparations.
- Published
- 2006