Design, construction, and characterization of high-performance membrane fusion devices with target-selectivity.
Membrane fusion proteins such as the hemagglutinin glycoprotein have target recognition and fusion accelerative domains, where some synergistically working elements are essential for target-selective and highly effective native membrane fusion systems. In this work, novel membrane fusion devices bearing such domains were designed and constructed. We selected a phenylboronic acid derivative as a recognition domain for a sugar-like target and a transmembrane-peptide (Leu-Ala sequence) domain interacting with the target membrane, forming a stable hydrophobic α-helix and accelerating the fusion process. Artificial membrane fusion behavior between the synthetic devices in which pilot and target liposomes were incorporated was characterized by lipid-mixing and inner-leaflet lipid-mixing assays. Consequently, the devices bearing both the recognition and transmembrane domains brought about a remarkable increase in the initial rate for the membrane fusion compared with the devices containing the recognition domain alone. In addition, a weakly acidic pH-responsive device was also constructed by replacing three Leu residues in the transmembrane-peptide domain by Glu residues. The presence of Glu residues made the acidic pH-dependent hydrophobic α-helix formation possible as expected. The target-selective liposome-liposome fusion was accelerated in a weakly acidic pH range when the Glu-substituted device was incorporated in pilot liposomes. The use of this pH-responsive device seems to be a potential strategy for novel applications in a liposome-based delivery system.
Department of Applied Molecular Chemistry, Graduate School of Industrial Technology, Nihon University, Narashino, Chiba 275-8575, Japan. kashiwada.ayumi@nihon-u.ac.jp
Ayumi
A
Kashiwada
Iori
I
Yamane
Mana
M
Tsuboi
Shun
S
Ando
Kiyomi
K
Matsuda
D000595:Amino Acid Sequence; D040761:Biomimetic Materials / Q000737:chemistry* / Q000378:metabolism*; D001897:Boronic Acids / Q000737:chemistry*; D002462:Cell Membrane / Q000378:metabolism; D015195:Drug Design; D006863:Hydrogen-Ion Concentration; D008081:Liposomes / Q000378:metabolism*; D008561:Membrane Fusion; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism; D013379:Substrate Specificity
D001897:Boronic Acids; D008081:Liposomes; D010455:Peptides; C010686:benzeneboronic acid