, Volume 6, Issue 4, pp 153–157 | Cite as

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

  • Madhavan Nallani
  • Mirjam Andreasson-Ochsner
  • Cherng-Wen Darren Tan
  • Eva-Kathrin Sinner
  • Yudi Wisantoso
  • Susana Geifman-Shochat
  • Walter Hunziker
Open Access


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.


  1. 1.
    Y. Shai, J. Membr. Biol. 182, 91 (2001).CrossRefGoogle Scholar
  2. 2.
    P. L. Yeagle and D. A. Lee, Biochim. Biophys. Acta 1565, 143 (2002).CrossRefGoogle Scholar
  3. 3.
    R. Robelek, E. S. Lemker, B. Wiltschi, V. Kirste, R. Naumann, D. Oesterhelt, and E. K. Sinner,. Angew. Chem Int. Ed. Engl. 46, 605 (2007).CrossRefGoogle Scholar
  4. 4.
    E. K. Sinner and W. Knoll, Curr. Opin. Chem. Biol. 5, 705 (2001).CrossRefGoogle Scholar
  5. 5.
    S. F. van Dongen, H. P. de Hoog, R. J. Peters, M. Nallani, R. J. Nolte, and J. C. van Hest, Chem Rev. 109, 6212 (2009).CrossRefGoogle Scholar
  6. 6.
    S. Belegrinou, J. Dorn, M. Kreiter, K. Kita-Tokarczyk, E. K. Sinner, and W. Meier, Soft Matter 6, 179 (2010).CrossRefGoogle Scholar
  7. 7.
    A. Graff, M. Sauer, G. P. Van, and W. Meier, Proc. Natl. Acad. Sci. U.S.A. 99, 5064 (2002).CrossRefGoogle Scholar
  8. 8.
    M. Kumar, M. Grzelakowski, J. Zilles, M. Clark, and W. Meier, Proc. Natl. Acad. Sci. U.S.A. 104, 20719 (2007).CrossRefGoogle Scholar
  9. 9.
    O. Onaca, P. Sarkar, D. Roccatano, T. Friedrich, B. Hauer, M. Grzelakowski, A. Guven, M. Fioroni, and U. Schwaneberg, Angew. Chem. Int. Ed. Engl. 47, 7029 (2008)CrossRefGoogle Scholar
  10. 10.
    M. Kaneda, S. M. Nomura, S. Ichinose, S. Kondo, K. Nakahama, K. Akiyoshi, and I. Morita, Biomaterials 30, 3971 (2009).CrossRefGoogle Scholar
  11. 11.
    H. Bermudez, A. K. Brannan, D. A. Hammer, F. S. Bates, and D. E. Discher, Macromolecules 35, 8203 (2002).CrossRefGoogle Scholar
  12. 12.
    K. Morita, M. Furuse, K. Fujimoto, and S. Tsukita, Proc. Natl. Acad. Sci. U.S.A. 96, 511 (1999).CrossRefGoogle Scholar
  13. 13.
    F. Ahmed and D. E. Discher, J. Controlled Release 96, 37 (2004).CrossRefGoogle Scholar
  14. 14.
    R. C. Macdonald, R. I. Macdonald, B. P. M. Menco, K. Takeshita, N. K. Subbarao, and L. R. Hu, Biochim. Biophys. Acta 1061, 297 (1991).CrossRefGoogle Scholar
  15. 15.
    M. Nirenberg and J. H. Matthaei, Proc. Nat. Acad. Sci. U.S.A. 47, 1588 (1961).CrossRefGoogle Scholar
  16. 16.
    F. Katzen, G. Chang, and W. Kudlicki, Trends Biotechnol. 23, 150 (2005).CrossRefGoogle Scholar
  17. 17.
    M. Furuse, K. Fujita, T. Hiiragi, K. Fujimoto, and S. Tsukita, J. Cell Biol. 141, 1539 (1998).CrossRefGoogle Scholar
  18. 18.
    A. Ranquin, W. Versees, W. Meier, J. Steyaert, and P. Van Gelder, Nano Lett. 5, 2220 (2005).CrossRefGoogle Scholar
  19. 19.
    R. Syeda, M. A. Holden, W. L. Hwang, and H. Bayley, J. Am. Chem. Soc. 130, 15543 (2008).CrossRefGoogle Scholar
  20. 20.
    W. Knoll, Ann. Rev. Phys. Chem. 49, 569 (1998)CrossRefGoogle Scholar
  21. 21.
    L. G. Fagerstam, A. Frostellkarlsson, R. Karlsson, B. Persson, and I. Ronnberg, J. Chromatogr. 597, 397 (1992).CrossRefGoogle Scholar
  22. 22.
    M. Malmqvist, Nature 361, 186 (1993)CrossRefGoogle Scholar
  23. 23.
    D. E. Discher and A. Eisenberg, Science 297, 967 (2002).CrossRefGoogle Scholar

Copyright information

© American Vacuum Society 2011

Authors and Affiliations

  • Madhavan Nallani
    • 1
  • Mirjam Andreasson-Ochsner
    • 1
  • Cherng-Wen Darren Tan
    • 1
  • Eva-Kathrin Sinner
    • 1
  • Yudi Wisantoso
    • 2
  • Susana Geifman-Shochat
    • 2
  • Walter Hunziker
    • 3
  1. 1.Institute of Materials Research and EngineeringA*STAR (Agency for Science, Technology and Research)Singapore
  2. 2.School of Biological SciencesNanyang Technological UniversitySingapore
  3. 3.Institute of Molecular and Cell BiologyA*STAR (Agency for Science, Technology and Research)Singapore

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