Low conductance gramicidin A channels are head-to-head dimers of beta 6.3-helices.

Weakly conductive, atypical channels were observed to form from highly purified Val1-gramicidin A in planar lipid bilayer membranes. The structure of these low-conductance channels (minis) was investigated by a detailed study of their channel forming characteristics. The possibility that minis originate from primary structural analogs or degradation products of gramicidin was considered and ruled out. In particular, spontaneous conductance changes in single channels demonstrated that minis can derive directly and reversibly from "standard" channels having the most common conductance level. The fraction of channels which are minis does not vary with changes in membrane gramicidin concentration, indicating that mini and standard channels have the same molecularity, that is, both are dimers. The mean lifetime of mini channels is only slightly shorter than that of standard channels, indicating that the six hydrogen bonds that stabilize the head-to-head dimer are minimally affected in minis. The fraction of channels which are minis is unaffected by the ionic strength, ionic composition, or pH of the bathing solution; it is also unaffected by the lipid composition of the bilayer. These findings are consistent with the hypothesis that minis arise from minor changes in the conformation of the Val1-gramicidin A molecule near the channel entrance or exit.