Brief Communication | Published:

A quantitative liposome microarray to systematically characterize protein-lipid interactions

Nature Methods volume 11, pages 4750 (2014) | Download Citation

Abstract

Lipids have a role in virtually all biological processes, acting as structural elements, scaffolds and signaling molecules, but they are still largely under-represented in known biological networks. Here we describe a liposome microarray–based assay (LiMA), a method that measures protein recruitment to membranes in a quantitative, automated, multiplexed and high-throughput manner.

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References

  1. 1.

    & Nature 443, 651–657 (2006).

  2. 2.

    et al. Mol. Syst. Biol. 6, 430–444 (2010).

  3. 3.

    , , , & Cell 144, 55–66 (2011).

  4. 4.

    & Methods 39, 122–133 (2006).

  5. 5.

    et al. Nat. Methods 9, 1189–1191 (2012).

  6. 6.

    , & J. Struct. Biol. 168, 90–94 (2009).

  7. 7.

    & Angew. Chem. Int. Edn Engl. 44, 6697–6700 (2005).

  8. 8.

    , , & Biophys. J. 91, 233–243 (2006).

  9. 9.

    , , & ChemBioChem 11, 848–865 (2010).

  10. 10.

    & Annu. Rev. Anal. Chem. (Palo Alto Calif.) 1, 801–832 (2008).

  11. 11.

    , , & J. Am. Chem. Soc. 131, 1810–1819 (2009).

  12. 12.

    , , & Anal. Chem. 70, 4974–4984 (1998).

  13. 13.

    et al. Nat. Methods 3, 385–390 (2006).

  14. 14.

    et al. Genome Biol. 7, R100 (2006).

  15. 15.

    Nat. Rev. Mol. Cell Biol. 9, 99–111 (2008).

  16. 16.

    et al. Nat. Rev. Mol. Cell Biol. 11, 103–112 (2010).

  17. 17.

    et al. Cell 152, 1008–1020 (2013).

  18. 18.

    & Proc. Natl. Acad. Sci. USA 107, 3436–3440 (2010).

  19. 19.

    et al. Nature 425, 686–691 (2003).

  20. 20.

    et al. Science 328, 593–599 (2010).

  21. 21.

    , , , & Nat. Biotechnol. 24, 79–88 (2006).

  22. 22.

    , , & Lab Chip 8, 274–279 (2008).

  23. 23.

    , & Nat. Protoc. 5, 491–502 (2010).

  24. 24.

    , & Lab Chip 5, 1393–1398 (2005).

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Acknowledgements

We dedicate this paper to T.P. We are grateful to C. Merten, M. Kaksonen, A. Galih, K. Kugler, C. Besir, M. Hsiung, M. Skruzny and the Protein Expression and Purification Core Facility for expert help and the sharing of reagents. We thank M. Mall (European Molecular Biology Laboratory) for the PKCγ and PKCδ (in the pEGFP-N3 vector). We also thank J.E.'s and other members of A.-C.G.'s groups for continuous discussions and support. This work was partially funded by the German Federal Ministry of Education and Research (BMBF; 01GS0865) in the framework of the IG-Cellular System genomics to A.-C.G. K.M. was supported by the Danish Natural Science Research Council (09-064986/FNU). A.-E.S. is supported by the European Molecular Biology Laboratory and the EU Marie Curie Actions Interdisciplinary Postdoctoral Cofunded Programme.

Author information

Author notes

    • Tony Pawson

    Deceased.

    • Antoine-Emmanuel Saliba
    •  & Ivana Vonkova

    These authors contributed equally to this work.

Affiliations

  1. Structural and Computational Biology Unit, European Molecular Biology Laboratory, Heidelberg, Germany.

    • Antoine-Emmanuel Saliba
    • , Ivana Vonkova
    • , Stefano Ceschia
    • , Kenji Maeda
    • , Samy Deghou
    • , Vera van Noort
    • , Peer Bork
    •  & Anne-Claude Gavin
  2. Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Toronto, Ontario, Canada.

    • Greg M Findlay
    •  & Tony Pawson
  3. Advanced Light Microscopy Facility, European Molecular Biology Laboratory, Heidelberg, Germany.

    • Christian Tischer
  4. Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany.

    • Jan Ellenberg

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Contributions

A.-E.S., I.V., J.E., V.v.N., P.B., T.P. and A.-C.G. designed the research, discussed the results and/or advised on the analyses; A.-E.S., I.V., S.C. and S.D. conducted the experiments and performed the analyses; G.M.F., K.M. and C.T. provided key technical expertise with instrumentation, protocols and reagents; and A.-E.S., I.V. and A.-C.G. wrote the manuscript with support from all the authors.

Competing interests

A.-E.S., I.V., J.E. and A.-C.G. are named on the international patent application PCT/EP2013/065256, "Giant liposome array for high-throughput lipid-protein interaction screening," which is based on the results described in this study.

Corresponding author

Correspondence to Anne-Claude Gavin.

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Figures 1–6, Supplementary Tables 1–3 and Supplementary Note

Videos

  1. 1.

    Bright-field image showing liposome formation on a thin agarose layer (TAL)

    At the initial time point (t = 0 s), a phosphate buffered saline (PBS, pH 7.4) is injected upon TAL inked with a lipid mixture composed of PC. Lipids self-assemble in liposomes in less than 3 minutes.

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DOI

https://doi.org/10.1038/nmeth.2734

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