A Modular Platform for One-Step Assembly of Multi-Component Membrane Systems by Fusion of Charged Proteoliposomes

Nat Commun. 2016 Oct 6;7:13025. doi: 10.1038/ncomms13025.

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 F1Fo ATP-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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / chemistry*
  • Calibration
  • Cell Membrane / enzymology
  • Cobalt / chemistry
  • Electron Transport
  • Escherichia coli / enzymology*
  • Fluoresceins / chemistry
  • Lipid Bilayers / chemistry
  • Liposomes / chemistry*
  • Membrane Fusion
  • Membrane Proteins / chemistry
  • Oxidoreductases / chemistry
  • Protein Binding
  • Proteolipids / chemistry*
  • Proton Pumps
  • Proton-Translocating ATPases / chemistry*
  • Soybeans / chemistry
  • Synthetic Biology

Substances

  • Fluoresceins
  • Lipid Bilayers
  • Liposomes
  • Membrane Proteins
  • Proteolipids
  • Proton Pumps
  • proteoliposomes
  • Cobalt
  • Adenosine Triphosphate
  • Oxidoreductases
  • Proton-Translocating ATPases
  • fluorexon