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Gating and conductance properties of a human delayed rectifier K+ channel expressed in frog oocytes.
Benndorf K, Koopmann R, Lorra C, Pongs O.
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1. The human delayed rectifier K+ channel h-DRK1, a homologue to the DRK1 channel in the rat, was expressed in Xenopus oocytes. Single-channel currents were measured in micropatches; macroscopic currents were measured either in macropatches, giant patches, or whole oocytes. 2. Macroscopic currents activated at -20 mV and more positive. The instantaneous current-voltage relationship rectified outwardly to a higher degree than predicted by the Goldman-Hodgkin-Katz equation. 3. With the giant patch technique, ionic and putative on- and off-gating currents were recorded simultaneously. The large ratio of the moved gating charges to the amplitude of the ionic current indicated that less than 1% of the gating channels actually opened. 4. The single-channel conductance between 0 and +80 mV was calculated to be 9.4 pS. The channels opened with sublevels which appeared either independently of the fully open level as separate openings, in conjunction with the opening to and closing from the fully open level, or by starting from and ending at the fully open level. 5. The channels opened with two voltage-independent open time constants in the range 1-10 ms (filter 1 kHz). The burst open probability was fitted monoexponentially with time constants in the range of tens of milliseconds. 6. Assuming a sequential Markovian model with four independent voltage-controlled transitions, fit of the steady-state open probability of macroscopic currents showed two components of activation differing in their half-maximal value. 7. The fit of time courses of cumulative first latency and ensemble-averaged currents in single-channel patches suggested that even a single channel may operate with the two different components of activation. 8. It is concluded that h-DRK1 channels considerably rectify in an outward direction and that an apparently flat voltage dependence of activation may be explained by the overlap of two different components.
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