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Journal for Biophysical Chemistry

Lipid-anchored DNA mediates vesicle fusion as observed by lipid and content mixing

Abstract

A general method for synthesizing 5- and 3-coupled DNA-lipid conjugates has been developed and employed in DNA-mediated vesicle fusion. Vesicles presenting complementary DNA fuse, resulting in both outer and inner leaflet mixing as well as content mixing. Fusion is maximized using 5′- and 3′-coupled DNA on opposite vesicle partners, rather than only 5′-coupled DNA, showing the importance of DNA orientation to the process. Lipid and content mixing assays show a dependence of fusion kinetics on the sequence and average number of DNA per vesicle. Vesicles without DNA or presenting noncomplementary sequences also appear to undergo some degree of lipid mixing or exchange, but no content mixing. Total lipid mixing appears to occur more efficiently than inner leaflet mixing and content mixing, and this may be explained by the observed nonspecific lipid mixing and/or the rise of a hemifused intermediate. The ability to control DNA sequence and the relative experimental simplicity of this system make it highly attractive to probe fundamental questions of membrane fusion using both ensemble and single vesicle assays.

References

  1. D. Papahadjopoulos, W. J. Vail, W. A. Pangborn, and G. Poste, Biochim. Biophys. Acta 448, 265 (1976).

    Article  CAS  Google Scholar 

  2. B. R. Lentz, G. F. McIntyre, D. J. Parks, J. C. Yates, and D. Massenburg, Biochemistry 31, 2643 (1992).

    Article  CAS  Google Scholar 

  3. Y. Gong, Y. Luo, and D. Bong, J. Am. Chem. Soc. 128, 14430 (2006).

    Article  CAS  Google Scholar 

  4. T. Weber, B. V. Zemelman, J. A. McNew, B. Westermann, M. Gmachl, F. Parlati, T. H. Söllner, and J. E. Rothman, Cell 92, 759 (1998).

    Article  CAS  Google Scholar 

  5. R. C. Lin and R. H. Scheller, Neuron 19, 1087 (1997).

    Article  CAS  Google Scholar 

  6. A. V. Pobbati, A. Stein, and D. Fasshauer, Science 313, 673 (2006).

    Article  CAS  Google Scholar 

  7. C. G. Schuette, K. Hatsuzawa, M. Margittai, A. Stein, D. Riedel, P. Küster, M. König, C. Seidel, and R. Jahn, Proc. Natl. Acad. Sci. U.S.A. 101, 2858 (2004).

    Article  CAS  Google Scholar 

  8. C. Yoshina-Ishii and S. G. Boxer, J. Am. Chem. Soc. 125, 3696 (2003).

    Article  CAS  Google Scholar 

  9. C. Yoshina-Ishii, G. P. Miller, M. L. Kraft, E. T. Kool, and S. G. Boxer, J. Am. Chem. Soc. 127, 1356 (2005).

    Article  CAS  Google Scholar 

  10. C. Yoshina-Ishii, Y.-H. M. Chan, J. M. Johnson, L. A. Kung, P. Lenz, and S. G. Boxer, Langmuir 22, 5682 (2006).

    Article  CAS  Google Scholar 

  11. Y.-H. M. Chan, P. Lenz, and S. G. Boxer, Proc. Natl. Acad. Sci. U.S.A. 104, 18913 (2007).

    Article  CAS  Google Scholar 

  12. G. Stengel, R. Zahn, and F. Höök, J. Am. Chem. Soc. 129, 9584 (2007).

    Article  CAS  Google Scholar 

  13. Y. Watanabe and M. Nakatomi, Tetrahedron 55, 9743 (1999).

    Article  CAS  Google Scholar 

  14. Y. Xu, S. A. Lee, T. G. Kutateladze, D. Sbrissa, A. Shisheva, and G. D. Prestwich, J. Am. Chem. Soc. 128, 885 (2006).

    Article  CAS  Google Scholar 

  15. M. Nazeem Nanjee, A. K. Gebre, and N. E. Miller, Clin. Chem. 37, 868 (1991).

    Google Scholar 

  16. J. Wilschut and D. Papahadjopoulos, Nature (London) 281, 690 (1979).

    Article  CAS  Google Scholar 

  17. Y. Xu, F. Zhang, Z. Su, J. A. McNew, and Y.-K. Shin, Nat. Struct. Mol. Biol. 12, 417 (2005).

    Article  CAS  Google Scholar 

  18. I. Pfeiffer and F. Höök, J. Am. Chem. Soc. 126, 10224 (2004).

    Article  CAS  Google Scholar 

  19. R. B. Sutton, D. Fasshauer, R. Jahn, and A. T. Brunger, Nature (London) 395, 347 (1998).

    Article  CAS  Google Scholar 

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Correspondence to Steven G. Boxer.

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Chan, YH.M., van Lengerich, B. & Boxer, S.G. Lipid-anchored DNA mediates vesicle fusion as observed by lipid and content mixing. Biointerphases 3, FA17–FA21 (2008). https://doi.org/10.1116/1.2889062

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  • DOI: https://doi.org/10.1116/1.2889062