title: Highly efficient macromolecule-sized poration of lipid bilayers by a synthetically evolved peptide.
abstract: Peptides that self-assemble, at low concentration, into bilayer-spanning pores which allow the passage of macromolecules would be beneficial in multiple areas of biotechnology. However, there are few, if any, natural or designed peptides that have this property. Here we show that the 26-residue peptide "MelP5", a synthetically evolved gain-of-function variant of the bee venom lytic peptide melittin identified in a high-throughput screen for small molecule leakage, enables the passage of macromolecules across bilayers under conditions where melittin and other pore-forming peptides do not. In surface-supported bilayers, MelP5 forms unusually high conductance, equilibrium pores at peptide:lipid ratios as low as 1:25000. The increase in bilayer conductance due to MelP5 is dramatically higher, per peptide, than the increase due to the parent sequence of melittin or other peptide pore formers. Here we also develop two novel assays for macromolecule leakage from vesicles, and we use them to characterize MelP5 pores in bilayers. We show that MelP5 allows the passage of macromolecules across vesicle membranes at peptide:lipid ratios as low as 1:500, and under conditions where neither osmotic lysis nor gross vesicle destabilization occur. The macromolecule-sized, equilibrium pores formed by MelP5 are unique as neither melittin nor other pore-forming peptides release macromolecules significantly under the same conditions. MelP5 thus appears to belong to a novel functional class of peptide that could form the foundation of multiple potential biotechnological applications.
authors.affiliation: Department of Biochemistry and Molecular Biology, Tulane University School of Medicine , New Orleans, Louisiana 70112, United States.
authors.forename: Gregory
authors.initials: G
authors.lastname: Wiedman
authors.forename: Taylor
authors.initials: T
authors.lastname: Fuselier
authors.forename: Jing
authors.initials: J
authors.lastname: He
authors.forename: Peter C
authors.initials: PC
authors.lastname: Searson
authors.forename: Kalina
authors.initials: K
authors.lastname: Hristova
authors.forename: William C
authors.initials: WC
authors.lastname: Wimley
mesh_terms: D000595:Amino Acid Sequence; D008051:Lipid Bilayers / Q000378:metabolism*; D008555:Melitten / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism*; D008969:Molecular Sequence Data; D016062:Porosity; D017433:Protein Structure, Secondary
chemical_list: D008051:Lipid Bilayers; D008555:Melitten