Skip to main content

Journal for Biophysical Chemistry

Biointerphases Cover Image

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

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. 1

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

    Article  CAS  Google Scholar 

  2. 2

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

    Article  CAS  Google 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).

    Article  CAS  Google Scholar 

  4. 4

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

    Article  CAS  Google 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).

    Article  Google Scholar 

  6. 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. 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. 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. 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. 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. 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. 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. 13

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

    Article  CAS  Google 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).

    Article  CAS  Google Scholar 

  15. 15

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

    Article  CAS  Google Scholar 

  16. 16

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

    Article  CAS  Google Scholar 

  17. 17

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

    Article  CAS  Google Scholar 

  18. 18

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

    Article  CAS  Google Scholar 

  19. 19

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

    Article  CAS  Google Scholar 

  20. 20

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

    Article  CAS  Google Scholar 

  21. 21

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

    Article  CAS  Google Scholar 

  22. 22

    M. Malmqvist, Nature 361, 186 (1993)

    Article  CAS  Google Scholar 

  23. 23

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

    Article  CAS  Google Scholar 

Download references

Author information

Affiliations

Authors

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Nallani, M., Andreasson-Ochsner, M., Tan, C.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

Download citation