title: A modular platform for one-step assembly of multi-component membrane systems by fusion of charged proteoliposomes.
abstract: An important goal in synthetic biology is the assembly of biomimetic cell-like structures, which combine multiple biological components in synthetic lipid vesicles. A key limiting assembly step is the incorporation of membrane proteins into the lipid bilayer of the vesicles. Here we present a simple method for delivery of membrane proteins into a lipid bilayer within 5 min. Fusogenic proteoliposomes, containing charged lipids and membrane proteins, fuse with oppositely charged bilayers, with no requirement for detergent or fusion-promoting proteins, and deliver large, fragile membrane protein complexes into the target bilayers. We demonstrate the feasibility of our method by assembling a minimal electron transport chain capable of adenosine triphosphate (ATP) synthesis, combining Escherichia coli F1FoATP-synthase and the primary proton pump bo3-oxidase, into synthetic lipid vesicles with sizes ranging from 100 nm to ∼10 μm. This provides a platform for the combination of multiple sets of membrane protein complexes into cell-like artificial structures.
authors.affiliation: Department of Physics, University of Oxford, Clarendon Laboratory, Parks Road, Oxford OX1 3PU, UK.
authors.forename: Robert R
authors.initials: RR
authors.lastname: Ishmukhametov
authors.affiliation: Department of Physics, University of Oxford, Clarendon Laboratory, Parks Road, Oxford OX1 3PU, UK.
authors.forename: Aidan N
authors.initials: AN
authors.lastname: Russell
authors.affiliation: Department of Physics, University of Oxford, Clarendon Laboratory, Parks Road, Oxford OX1 3PU, UK.
authors.forename: Richard M
authors.initials: RM
authors.lastname: Berry
mesh_terms: D000255:Adenosine Triphosphate / Q000737:chemistry*; D002138:Calibration; D002462:Cell Membrane / Q000201:enzymology; D003035:Cobalt / Q000737:chemistry; D004579:Electron Transport; D004926:Escherichia coli / Q000201:enzymology*; D005452:Fluoresceins / Q000737:chemistry; D008051:Lipid Bilayers / Q000737:chemistry; D008081:Liposomes / Q000737:chemistry*; D008561:Membrane Fusion; D008565:Membrane Proteins / Q000737:chemistry; D010088:Oxidoreductases / Q000737:chemistry; D011485:Protein Binding; D011510:Proteolipids / Q000737:chemistry*; D017494:Proton Pumps; D006180:Proton-Translocating ATPases / Q000737:chemistry*; D013025:Glycine max / Q000737:chemistry; D058615:Synthetic Biology
chemical_list: D005452:Fluoresceins; D008051:Lipid Bilayers; D008081:Liposomes; D008565:Membrane Proteins; D011510:Proteolipids; D017494:Proton Pumps; C020485:proteoliposomes; D003035:Cobalt; D000255:Adenosine Triphosphate; D010088:Oxidoreductases; D006180:Proton-Translocating ATPases; C007740:fluorexon