Delivery of membrane proteins into small and giant unilamellar vesicles by charge-mediated fusion.
One of the current challenges in synthetic biology is the production of stable membrane mimetic systems and the insertion of components in these systems. Here, we employ fusion of oppositely charged liposomes to deliver separately reconstituted membrane proteins into a common lipid bilayer. After a systematic evaluation of different lipid compositions by lipid mixing and size distribution analysis, suitable conditions were further investigated for proteoliposome fusion. With this technique, we functionally coreconstituted bo3 oxidase and ATP synthase from Escherichia coli into unilamellar liposomes ranging from 100 nm to 50 μm in size. The presented method is a simple and versatile tool for oriented membrane protein reconstitution to produce biomimetic systems with increased complexity.
Department of Chemistry and Biochemistry, University of Bern, Switzerland.|Graduate School for Cellular and Biomedical Sciences, University of Bern, Switzerland.
Olivier
O
Biner
Department of Chemistry and Biochemistry, University of Bern, Switzerland.|Graduate School for Cellular and Biomedical Sciences, University of Bern, Switzerland.
Thomas
T
Schick
Department of Chemistry and Biochemistry, University of Bern, Switzerland.
Yannic
Y
Müller
Department of Chemistry and Biochemistry, University of Bern, Switzerland.
Christoph
0000-0002-4642-6088
C
von Ballmoos
D025181:ATP Synthetase Complexes / Q000737:chemistry*; D040761:Biomimetic Materials / Q000737:chemistry; D004926:Escherichia coli / Q000201:enzymology*; D029968:Escherichia coli Proteins / Q000737:chemistry*; D008565:Membrane Proteins / Q000737:chemistry*; D053835:Unilamellar Liposomes / Q000737:chemistry*
D029968:Escherichia coli Proteins; D008565:Membrane Proteins; D053835:Unilamellar Liposomes; D025181:ATP Synthetase Complexes
charged lipids; giant unilamellar vesicle; liposomes; membrane fusion; protein reconstitution; respiratory chain enzymes