Design and Synthesis of Amphiphilic α-Helical Model Peptides with Systematically Varied Hydrophobic−Hydrophilic Balance and Their Interaction with Lipid- and Bio-Membranes

Abstract

Five amphiphilic α-helical peptides of 18 residues containing a hydrophobic Trp residue as a fluorescence probe were designed. The peptides were made up of hydrophobic Leu and hydrophilic Lys residues of a ratio of 13:5, 11:7, 9:9, 7:11, and 5:13 (abbreviated as Hels 13−5, 11−7, 9−9, 7−11, and 5−13, respectively). These peptides generate ideal amphiphilic α-helical structures, which have systematically varied hydrophobic−hydrophilic balance (relative amphiphilic potential) as a result of different hydrophobicities and almost the same hydrophobic moments. Their hydrophobic−hydrophilic balance was estimated both theoretically from the calculated hydrophobicity values (or the magnitude of hydrophobic faces) and experimentally from the retention times in reverse phase high-performance liquid chromatography (RP-HPLC). Circular dichroism, liposome-lytic, and Trp-fluorescent studies in buffer and in the presence of acidic and neutral liposomes clearly showed that the increasing hydrophobic face area not only increases the affinity for lipid but also increases the trend of self-association. The structure−activity relationship estimated by means of leakage ability and hemolytic activity demonstrated that the model- and bio-membrane perturbation ability is completely parallel to the magnitude of the hydrophobic face area. The lipid-binding study in guanidine hydrochloride solution showed that the peptides with a hydrophobic face larger than the hydrophilic face (Hels 13−5 and 11−7) immerse their hydrophobic regions in lipid bilayers and that the inverse ones (Hels 7−11 and 5−13) interact only between the anionic lipid head groups and cationic peptide residues on liposome surfaces. The peptide Hel 9−9, which has exactly the same hydrophobic and hydrophilic regions, was found to be at a critical boundary among these peptides in terms of (1) behavior of peptide self-aggregation in buffer solution and membrane perturbation ability, (2) transfer from bulk solution to neutral lipid bilayers, and (3) necessity of charge interaction in lipid−peptide binding.