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Biointerphases

Journal for Biophysical Chemistry

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

Biointerphases volume 6, pages 153–157 (2011)Cite this article

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.

References

  1. Y. Shai, J. Membr. Biol. 182, 91 (2001).

    Article  CAS  Google Scholar 

  2. P. L. Yeagle and D. A. Lee, Biochim. Biophys. Acta 1565, 143 (2002).

    Article  CAS  Google Scholar 

  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).

    Article  CAS  Google Scholar 

  4. E. K. Sinner and W. Knoll, Curr. Opin. Chem. Biol. 5, 705 (2001).

    Article  CAS  Google Scholar 

  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).

    Article  Google Scholar 

  6. S. Belegrinou, J. Dorn, M. Kreiter, K. Kita-Tokarczyk, E. K. Sinner, and W. Meier, Soft Matter 6, 179 (2010).

    Article  CAS  Google Scholar 

  7. A. Graff, M. Sauer, G. P. Van, and W. Meier, Proc. Natl. Acad. Sci. U.S.A. 99, 5064 (2002).

    Article  CAS  Google Scholar 

  8. M. Kumar, M. Grzelakowski, J. Zilles, M. Clark, and W. Meier, Proc. Natl. Acad. Sci. U.S.A. 104, 20719 (2007).

    Article  CAS  Google Scholar 

  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)

    Article  CAS  Google Scholar 

  10. M. Kaneda, S. M. Nomura, S. Ichinose, S. Kondo, K. Nakahama, K. Akiyoshi, and I. Morita, Biomaterials 30, 3971 (2009).

    Article  CAS  Google Scholar 

  11. H. Bermudez, A. K. Brannan, D. A. Hammer, F. S. Bates, and D. E. Discher, Macromolecules 35, 8203 (2002).

    Article  CAS  Google Scholar 

  12. K. Morita, M. Furuse, K. Fujimoto, and S. Tsukita, Proc. Natl. Acad. Sci. U.S.A. 96, 511 (1999).

    Article  CAS  Google Scholar 

  13. F. Ahmed and D. E. Discher, J. Controlled Release 96, 37 (2004).

    Article  CAS  Google Scholar 

  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).

    Article  CAS  Google Scholar 

  15. M. Nirenberg and J. H. Matthaei, Proc. Nat. Acad. Sci. U.S.A. 47, 1588 (1961).

    Article  CAS  Google Scholar 

  16. F. Katzen, G. Chang, and W. Kudlicki, Trends Biotechnol. 23, 150 (2005).

    Article  CAS  Google Scholar 

  17. M. Furuse, K. Fujita, T. Hiiragi, K. Fujimoto, and S. Tsukita, J. Cell Biol. 141, 1539 (1998).

    Article  CAS  Google Scholar 

  18. A. Ranquin, W. Versees, W. Meier, J. Steyaert, and P. Van Gelder, Nano Lett. 5, 2220 (2005).

    Article  CAS  Google Scholar 

  19. R. Syeda, M. A. Holden, W. L. Hwang, and H. Bayley, J. Am. Chem. Soc. 130, 15543 (2008).

    Article  CAS  Google Scholar 

  20. W. Knoll, Ann. Rev. Phys. Chem. 49, 569 (1998)

    Article  CAS  Google Scholar 

  21. L. G. Fagerstam, A. Frostellkarlsson, R. Karlsson, B. Persson, and I. Ronnberg, J. Chromatogr. 597, 397 (1992).

    Article  CAS  Google Scholar 

  22. M. Malmqvist, Nature 361, 186 (1993)

    Article  CAS  Google Scholar 

  23. D. E. Discher and A. Eisenberg, Science 297, 967 (2002).

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Institute of Materials Research and Engineering, A*STAR (Agency for Science, Technology and Research), 3 Research Link, Singapore, 117602

    Madhavan Nallani, Mirjam Andreasson-Ochsner, Cherng-Wen Darren Tan & Eva-Kathrin Sinner

  2. School of Biological Sciences, Nanyang Technological University, 60 Nanyang Avenue, Singapore, 637551

    Yudi Wisantoso & Susana Geifman-Shochat

  3. Institute of Molecular and Cell Biology, A*STAR (Agency for Science, Technology and Research), 61 Biopolis Dr, Singapore, 138673

    Walter Hunziker

Authors
  1. Madhavan Nallani
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  2. Mirjam Andreasson-Ochsner
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  3. Cherng-Wen Darren Tan
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  4. Eva-Kathrin Sinner
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  5. Yudi Wisantoso
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  6. Susana Geifman-Shochat
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  7. Walter Hunziker
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Nallani, M., Andreasson-Ochsner, M., Tan, CW.D. et al. Proteopolymersomes: In vitro production of a membrane protein in polymersome membranes. Biointerphases 6, 153–157 (2011). https://doi.org/10.1116/1.3644384

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  • DOI: https://doi.org/10.1116/1.3644384