Published online March 31, 2008
doi:10.1085/jgp.200809956
The Journal of General Physiology, Vol. 131, No. 4, 365-378
The Rockefeller University Press, 0022-1295 $30.00
© 2008 Schroeder et al.
Tl+-induced µs Gating of Current Indicates Instability of the MaxiK Selectivity Filter as Caused by Ion/Pore Interaction
Indra Schroeder and
Ulf-Peter Hansen
Department of Structural Biology, University of Kiel, 24098 Kiel, Germany
Correspondence to Ulf-Peter Hansen: uphansen{at}zbm.uni-kiel.de
Patch clamp experiments on single MaxiK channels expressed in HEK293 cells were performed at high temporal resolution (50-kHz filter) in asymmetrical solutions containing 0, 25, 50, or 150 mM Tl+ on the luminal or cytosolic side with [K+] + [Tl+] = 150 mM and 150 mM K+ on the other side. Outward current in the presence of cytosolic Tl+ did not show fast gating behavior that was significantly different from that in the absence of Tl+. With luminal Tl+ and at membrane potentials more negative than –40 mV, the single-channel current showed a negative slope resistance concomitantly with a flickery block, resulting in an artificially reduced apparent single-channel current Iapp. The analysis of the amplitude histograms by β distributions enabled the estimation of the true single-channel current and the determination of the rate constants of a simple two-state O-C Markov model for the gating in the bursts. The voltage dependence of the gating ratio R = Itrue/Iapp = (kCO + kOC)/kCO could be described by exponential functions with different characteristic voltages above or below 50 mM Tl+. The true single-channel current Itrue decreased with Tl+ concentrations up to 50 mM and stayed constant thereafter. Different models were considered. The most likely ones related the exponential increase of the gating ratio to ion depletion at the luminal side of the selectivity filter, whereas the influence of [Tl+] on the characteristic voltage of these exponential functions and of the value of Itrue were determined by [Tl+] at the inner side of the selectivity filter or in the cavity.
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