Permeation characteristics of gramicidin conformers.

To investigate the molecular origin of decreased conductance in variant gramicidin channels, we examined the current-voltage (IV) characteristics of single Val1-gramicidin A channels. Unlike standard channels, all variant channels showed pronounced rectification even though bathing solutions were symmetrical. Moreover, channels of lower conductance consistently showed more pronounced rectification. Analysis within the framework of a three-barrier, two-site, single-filing model indicates that the shape of the variant channel IVs could be best explained by an increase in binding affinity near one of the two channel entrances. This conclusion was further tested by characterizing single channel IVs in bi-ionic solutions having different cationic species at each channel entrance. In Cs/Na bi-ionic solutions, reversal potentials of variant channels often differed by a small but significant amount from those of standard channels. When a membrane potential was applied, the ionic currents tended to be reduced more when flowing from the Na+ side than the Cs+ side. These observations support the conclusion that variant channels have increased binding affinity at one end of the channel. Furthermore, H+ currents were increased while Ag+ currents were unaltered for most variant channels exhibiting decreased Na+ or Cs+ currents. The increased H+ conductance argues against long-range coulombic forces as the basis for decreased Na+ or Cs+ conductance while the normal Ag+ conductance suggests that the binding site field strength increases by a change in carbonyl geometry at the channel entrance.