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A quantitative liposome microarray to systematically characterize protein-lipid interactions

Nature Methods volume 11, pages 47–50 (2014)Cite this article

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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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Figure 1: Liposome microarray–based assay (LiMA) experimental principle and overview.
Figure 2: LiMA quality and validation.
Figure 3: An E108K mutation in the SOS1 histone-fold domain (SOS-HF) affects its affinity for lipids.

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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.

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Author notes

  1. Tony Pawson: Deceased.

  2. Antoine-Emmanuel Saliba and Ivana Vonkova: These authors contributed equally to this work.

Authors and 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

Authors
  1. Antoine-Emmanuel Saliba
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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.

Corresponding author

Correspondence to Anne-Claude Gavin.

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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.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–6, Supplementary Tables 1–3 and Supplementary Note (PDF 1596 kb)

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. (MOV 1082 kb)

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Saliba, AE., Vonkova, I., Ceschia, S. et al. A quantitative liposome microarray to systematically characterize protein-lipid interactions. Nat Methods 11, 47–50 (2014). https://doi.org/10.1038/nmeth.2734

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