Self-assembly of designed antimicrobial peptides in solution and micelles.
Hydrophobic interactions are responsible for stabilizing leucine zippers in peptides containing heptad repeats. The effects of substituting leucine by phenylalanine and alanine by glycine on the self-assembly of coiled-coils were examined in minimalist antimicrobial peptides designed to form amphipathic alpha-helices. The secondary structure of these peptides was monitored in solution and in diphosphocholine (DPC) micelles using circular dichroism spectroscopy. The leucine peptides (KLAKLAK)3 and (KLAKKLA)n (n = 3, 4) become alpha-helical with increasing concentrations of salt, peptide, and DPC. The aggregation state and equilibrium constant for self-association of the peptides were measured by sedimentation equilibrium. The glycine peptide (KLGKKLG)3 does not self-associate. The leucine peptides and phenylalanine peptides (KFAKFAK)3 and (KFAKKFA)n (n = 3, 4) are in a monomer-tetramer equilibrium in solution, with the phenylalanine zippers being 2-4 kcal/mol less stable than the equivalent leucine zippers. Thermodynamic parameters for the association reaction were calculated from the temperature dependence of the association constants. Leucine zipper formation has DeltaCp = 0, whereas phenylalanine zipper formation has a small negative DeltaCp, presumably due to the removal of the larger surface area of phenylalanine from water. Self-association of the peptides is coupled to formation of a hydrophobic core as detected using 1-anilino-naphthalene-8-sulfonate fluorescence. Carboxyfluorescein-labeled peptides were used to determine the aggregation state of (KLAKKLA)3 and (KLGKKLG)3 in DPC micelles. (KLAKKLA)3 forms dimers, and (KLGKKLG)3 is a monomer. Aggregation appears to correlate with the cytotoxicity of these peptides.
Javadpour
M M
MM
Department of Chemistry, Louisiana State University, Baton Rouge, Louisiana 70803, USA.
Barkley
M D
MD
D016475:3T3 Cells; D000817:Anilino Naphthalenesulfonates; D000818:Animals; D000900:Anti-Bacterial Agents / Q000737:chemistry* / Q000494:pharmacology; D002942:Circular Dichroism; D005452:Fluoresceins; D005456:Fluorescent Dyes; D006090:Gram-Negative Bacteria / Q000187:drug effects; D006094:Gram-Positive Bacteria / Q000187:drug effects; D016350:Leucine Zippers; D051379:Mice; D008823:Micelles; D010455:Peptides; D010767:Phosphorylcholine / Q000031:analogs & derivatives; D017433:Protein Structure, Secondary; D012996:Solutions; D013816:Thermodynamics
D000817:Anilino Naphthalenesulfonates; D000900:Anti-Bacterial Agents; D005452:Fluoresceins; D005456:Fluorescent Dyes; D008823:Micelles; D010455:Peptides; D012996:Solutions; D010767:Phosphorylcholine; C024098:6-carboxyfluorescein; C028810:dodecylphosphocholine; C027132:1-anilino-8-naphthalenesulfonate