Biointerphases

Journal for Biophysical Chemistry

Biointerphases Cover Image
Open Access

Proteopolymersomes: In vitro production of a membrane protein in polymersome membranes

  • Madhavan Nallani1,
  • Mirjam Andreasson-Ochsner1,
  • Cherng-Wen Darren Tan1,
  • Eva-Kathrin Sinner1,
  • Yudi Wisantoso2,
  • Susana Geifman-Shochat2 and
  • Walter Hunziker3
Biointerphases6:2

https://doi.org/10.1116/1.3644384

Received: 18 July 2011

Accepted: 9 September 2011

Abstract

Polymersomes are stable self-assembled architectures which mimic cell membranes. For characterization, membrane proteins can be incorporated into such bio-mimetic membranes by reconstitution methods, leading to so-called proteopolymersomes. In this work, we demonstrate the direct incorporation of a membrane protein into polymersome membranes by a cell-free expression system. Firstly, we demonstrate pore formation in the preformed polymersome membrane using α-hemolysin. Secondly, we use claudin-2, a protein involved in cell-cell interactions, to demonstrate the in vitro expression of a membrane protein into these polymersomes. Surface plasmon resonance (Biacore) binding studies with the claudin-2 proteopolymersomes and claudin-2 specific antibodies are performed to show the presence of the in vitro expressed protein in polymersome membranes.