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Updated: Aug 15, 2026

Examining the Conformational Dynamics of Membrane Proteins in situ with Site-directed Fluorescence Labeling
Published on: May 29, 2011
Characterization of a cation channel on the apical surface of the frog lens epithelium
Abstract:
The properties of a single conductance pathway of the apical (fibre-facing) surface of the frog lens epithelium are reported. Using the patch-clamp technique (Hamill, Marty, Neher, Sakmann & Sigworth, 1981), the most common single-channel currents had an amplitude of 1.9 pA, the mean open time 2.1 ms and a conductance of 25 pS. One open-state time constant (to = 3.3 ms) and two closed-state time constants (tau c1 = 0.9 ms, tau c2 = 23.1 ms) were resolved. The channel current and the mean open time were both increased when Ca2+ was removed from the external solution and the open time distribution was no longer fitted by a single exponential. Multiple-channel events in cell-attached patches containing two or more identical channels were distributed in a binomial fashion and the probability that an individual channel was open, obtained by fitting the binomial distribution, was 0.039. The channel was found to have a Na+:K+ selectivity ratio of 3:1. When Ca2+ was removed from the pipette solution the probability that an individual channel was open increased to 0.137 and the Na+:K+ selectivity ratio increased to 4:1. Channel activity was observed in the presence of tetrodotoxin (10(-6) M) in the bathing medium and the pipette solution but was abolished by internal perfusion of the patch pipette with 0.5 x 10(-4) M amiloride. this apical conductance pathway is identified as an amiloride-sensitive cation channel. These channels are clustered in groups on the apical membrane, spontaneously active at the resting potential and with the possibility of altering their Na+:K+ selectivity. They represent a distinct type of channel, that differ from nerve and muscle Na+ channels in their manner of activation, but do share some common features with both Na+ and Ca2+ channels in excitable cells.
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