Formamidinium-induced dimer stabilization and flicker block behavior in homo- and heterodimer channels formed by gramicidin A and N-acetyl gramicidin A.
Compared to the N-formyl gramicidin A (GA), the N-acetyl gramicidin A (NAG) channel has unchanged conductance in 1 M NH4+ (gamma NN/gamma GG = 1, conductance ratio) but reduced conductance in 1 M K+ (gamma NN/gamma GG = 0.6) methylammonium (gamma NN/gamma GG = 0.3), and formamidinium (gamma NN/gamma GG = 0.1) solutions. Except with formamidinium, "flicker blocks" are evident even at low cutoff frequencies. For all cations studied, channel lifetimes of N-acetyl homodimers (NN) are approximately 50-fold shorter than those of the GA homodimer (GG). The novel properties of GA channels in formamidinium solution (supralinear current-voltage relations and dimer stabilization (Seoh and Busath, 1993)) also appear in NN channels. The average single channel lifetime in 1 M formamidinium solution at 100 mV is 6-7-fold longer than in K+ and methylammonium solutions and, like in the GA channel, significantly decreases with increasing membrane potential. Experiments with mixtures of the two peptides, GA and NAG, showed three main conductance peaks. Oriented hybrids were formed utilizing the principle that monomers remain in one leaflet of the bilayer (O'Connell et al., 1990). With GA at the polarized side and NAG at the grounded side, at positive potentials (in which case hybrids were designated GN) and at negative potentials (in which case hybrids were designated NG), channels had the same conductances and channel properties at all potentials studied. Flicker blocks were not evident in the hybrid channels, which suggests that both N-acetyl methyl groups at the junction of the dimer are required to cause flickers. Channel lifetimes in hybrids are only approximately threefold shorter than those of the GG channels, and channel conductances are similar to those of GG rather than NN channels. We suggest that acetyl-acetyl crowding at the dimeric junction in NN channels cause dimer destabilization, flickers, and increased selectivity in N-acetyl gramicidin channels.
Seoh
S A
SA
Section of Physiology, Brown University, Providence, Rhode Island 02912.
Busath
D D
DD
D000578:Amidines / Q000494:pharmacology; D055592:Biophysical Phenomena; D001703:Biophysics; D004553:Electric Conductivity; D006096:Gramicidin / Q000737:chemistry* / Q000031:analogs & derivatives; D006860:Hydrogen Bonding; D007473:Ion Channels / Q000737:chemistry* / Q000187:drug effects; D008051:Lipid Bilayers / Q000737:chemistry; D008564:Membrane Potentials; D008744:Methylamines / Q000494:pharmacology; D008958:Models, Molecular; D011188:Potassium / Q000494:pharmacology; D011487:Protein Conformation / Q000187:drug effects; D000644:Quaternary Ammonium Compounds / Q000494:pharmacology
D000578:Amidines; D007473:Ion Channels; D008051:Lipid Bilayers; D008744:Methylamines; D000644:Quaternary Ammonium Compounds; D006096:Gramicidin; C077922:formamidine; C032275:N-acetylgramicidin; C027451:methylamine; D011188:Potassium