Amino acid sequence modulation of gramicidin channel function: effects of tryptophan-to-phenylalanine substitutions on the single-channel conductance and duration.
Linear gramicidins with one, two, or three Trp----Phe substitutions in the gramicidin A sequence form beta 6.3-helical channels that have widely varying conductances and average durations. The variations in single-channel conductance and average duration are uncoupled. The single-channel conductance decreases as a monotonic function of the number of Trp----Phe substitutions, and the relative conductance decrease induced by a given Trp----Phe substitution is only weakly affected by substitutions at other positions. These results suggest that each Trp influences the conductance independently, most likely through electrostatic interactions between the Trp dipole(s) and the permeant ion (as was deduced previously for aromatic side-chain substitutions at position one [Koeppe, R. E., Mazet, J.-L., & Andersen, O. S. (1990) Biochemistry 29 (2), 512-520]). Trp----Phe substitutions exert a complex, nonadditive influence on average duration as well as the energetics of heterodimer formation. These changes are presumably due to sequence-specific differences in the channel's surface chemistry--which may be related to ability of the Trp indole NH moieties to form hydrogen bonds with the lipid backbone oxygens and/or interfacial H2O.
Becker
M D
MD
Department of Physiology and Biophysics, Cornell University Medical College, New York 10021.
Greathouse
D V
DV
Koeppe
R E
RE
Andersen
O S
OS
D000595:Amino Acid Sequence; D006096:Gramicidin / Q000737:chemistry*; D006860:Hydrogen Bonding; D007473:Ion Channels / Q000737:chemistry* / Q000187:drug effects / Q000502:physiology; D008051:Lipid Bilayers / Q000737:chemistry; D008564:Membrane Potentials / Q000187:drug effects*; D008969:Molecular Sequence Data; D010649:Phenylalanine / Q000737:chemistry* / Q000494:pharmacology; D011487:Protein Conformation; D013329:Structure-Activity Relationship; D013816:Thermodynamics; D014364:Tryptophan / Q000737:chemistry* / Q000494:pharmacology
D007473:Ion Channels; D008051:Lipid Bilayers; D006096:Gramicidin; D010649:Phenylalanine; D014364:Tryptophan