Delivery of membrane proteins into small and giant unilamellar vesicles by charge-mediated fusion

Biner, Olivier, Thomas Schick, Yannic Müller, and Christoph Ballmoos. FEBS letters 590, no. 14 (2016): 2051-2062.

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 co-reconstituted 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.

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