title: Construction of a pH-responsive artificial membrane fusion system by using designed coiled-coil polypeptides.
abstract: In many viruses, pH-responsive coiled-coil domains in the specific fusion proteins play important roles in membrane fusion and the infection of viruses into host cells. To investigate the relationship between the conformational change of the coiled coil and the fusion process, we have introduced a de novo designed polypeptide as a model system of the coiled-coil domain. This system enables the systematic study of the dynamics of pH-responsive coiled-coil polypeptide-membrane interactions. First, we designed and synthesized pH-responsive isoleucine-zipper triple-stranded coiled-coil polypeptides. Then the relationship between the pH-induced conformational change of the polypeptide and the membrane's interactive properties was studied by physicochemical methods. Structural changes in the designed polypeptides were examined by means of circular dichroism measurements. And finally, the behavior of the membrane fusion was investigated by leakage of liposomal contents, turbidity analysis, dynamic light scattering, and lipid mixing experiments. Our data show that coiled-coil formation under acidic pH conditions enhances polypeptide-induced membrane fusion. The results in this study demonstrate that an artificial membrane fusion system can be constructed on a molecular level by the use of a pH-responsive isoleucine-zipper triple-stranded coiled-coil polypeptide.
authors.affiliation: Department of Applied Molecular Chemistry, College of Industrial Technology, Nihon University, 1-2-1 Izumi-cho, Narashino, Chiba, Japan. a5kasiwa@cit.nihon-u.ac.jp
authors.forename: Ayumi
authors.initials: A
authors.lastname: Kashiwada
authors.forename: Kiyomi
authors.initials: K
authors.lastname: Matsuda
authors.forename: Toshihisa
authors.initials: T
authors.lastname: Mizuno
authors.forename: Toshiki
authors.initials: T
authors.lastname: Tanaka
mesh_terms: D002942:Circular Dichroism; D031541:Fluorescence Resonance Energy Transfer; D006863:Hydrogen-Ion Concentration; D008081:Liposomes; D008561:Membrane Fusion; D008567:Membranes, Artificial; D010455:Peptides / Q000737:chemistry*; D011487:Protein Conformation
chemical_list: D008081:Liposomes; D008567:Membranes, Artificial; D010455:Peptides