C-terminally shortened alamethicin on templates: influence of the linkers on conductances.
In order to test the influence of chemical modifications designed to allow covalent coupling of channel-forming peptide motifs into variable sized oligomers, a series of alamethicin derivatives was prepared. The building block encompassing the N-terminal 1-17 residues of alamethicin behaved normally in the conductance assay on planar lipid bilayers, albeit at higher concentration and with a slightly reduced voltage-dependence. A linker Ac-K-OCH(2)C(6)H(4)CH(3)p attached via the epsilon amino group of lysine to the C-terminus of alamethicin(1-17) increased membrane affinity. The latter was further enhanced in a dimer and a tetramer in which alamethicin(1-17) chains were tethered to di- or tetra-lysine linkers, respectively, but macroscopic current-voltage curves displayed much reduced voltage-dependencies and reversed hysteresis. An usual behaviour with high voltage-dependence was restored with the modified dimer of alamethicin(1-17) in which alanine separated the two consecutive lysine residues in the linker. Of special interest was the development of a 'negative resistance' branch in macroscopic current-voltage curves for low concentrations of this dimer with the more flexible linker. Single channel events displayed only one single open state with fast kinetics and whose conductance matches that of the alamethicin heptamer or octamer.
Duclohier
H
H
UMR 6522 CNRS-Université de Rouen, Institut Fédératif de Recherches Multidisciplinaires sur les Peptides (IFRMP 23), 76821, Mont-Saint-Aignan, Cedex, France. herve.duclohier@univ-rouen.fr
Kociolek
K
K
Stasiak
M
M
Leplawy
M T
MT
Marshall
G R
GR
D000408:Alamethicin / Q000031:analogs & derivatives* / Q000737:chemistry*; D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000737:chemistry*; D001665:Binding Sites; D019281:Dimerization; D004553:Electric Conductivity; D066298:In Vitro Techniques; D007700:Kinetics; D008051:Lipid Bilayers / Q000737:chemistry; D008239:Lysine / Q000737:chemistry; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry*; D011487:Protein Conformation
D000900:Anti-Bacterial Agents; D008051:Lipid Bilayers; D010446:Peptide Fragments; D000408:Alamethicin; D008239:Lysine