In Vivo Profiling and Visualization of Cellular Protein–Lipid Interactions Using Bifunctional Fatty Acids

Authors

  • Dr. Per Haberkant,

    Corresponding author
    1. European Molecular Biology Laboratory, Meyerhofstrasse 1, 69117 Heidelberg (Germany)
    2. Membrane Biochemistry & Biophysics, Biomolecular Mass Spectrometry & Proteomics Group, Developmental Biology, Bijvoet Center for Biomolecular Research, Utrecht University, 3584 CH Utrecht (The Netherlands)
    • European Molecular Biology Laboratory, Meyerhofstrasse 1, 69117 Heidelberg (Germany)
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  • Dr. Reinout Raijmakers,

    1. Membrane Biochemistry & Biophysics, Biomolecular Mass Spectrometry & Proteomics Group, Developmental Biology, Bijvoet Center for Biomolecular Research, Utrecht University, 3584 CH Utrecht (The Netherlands)
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  • Dr. Marjolein Wildwater,

    1. Membrane Biochemistry & Biophysics, Biomolecular Mass Spectrometry & Proteomics Group, Developmental Biology, Bijvoet Center for Biomolecular Research, Utrecht University, 3584 CH Utrecht (The Netherlands)
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  • Timo Sachsenheimer,

    1. Heidelberg University Biochemistry Center, Im Neuenheimer Feld 328, 69120 Heidelberg (Germany)
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  • Priv.-Doz. Dr. Britta Brügger,

    1. Heidelberg University Biochemistry Center, Im Neuenheimer Feld 328, 69120 Heidelberg (Germany)
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  • Dr. Kenji Maeda,

    1. European Molecular Biology Laboratory, Meyerhofstrasse 1, 69117 Heidelberg (Germany)
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  • Dr. Martin Houweling,

    1. Department of Biochemistry and Cell Biology, Faculty of Veterinary Medicine, P.O. Box 80176, 3508 TD Utrecht (The Netherlands)
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  • Dr. Anne-Claude Gavin,

    1. European Molecular Biology Laboratory, Meyerhofstrasse 1, 69117 Heidelberg (Germany)
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  • Priv.-Doz. Dr. Carsten Schultz,

    1. European Molecular Biology Laboratory, Meyerhofstrasse 1, 69117 Heidelberg (Germany)
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  • Prof. Gerrit van Meer,

    1. Membrane Biochemistry & Biophysics, Biomolecular Mass Spectrometry & Proteomics Group, Developmental Biology, Bijvoet Center for Biomolecular Research, Utrecht University, 3584 CH Utrecht (The Netherlands)
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  • Prof. Albert J. R. Heck,

    1. Membrane Biochemistry & Biophysics, Biomolecular Mass Spectrometry & Proteomics Group, Developmental Biology, Bijvoet Center for Biomolecular Research, Utrecht University, 3584 CH Utrecht (The Netherlands)
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  • Prof. Joost C. M. Holthuis

    1. Membrane Biochemistry & Biophysics, Biomolecular Mass Spectrometry & Proteomics Group, Developmental Biology, Bijvoet Center for Biomolecular Research, Utrecht University, 3584 CH Utrecht (The Netherlands)
    2. Molecular Cell Biology, University of Osnabrück, Barbarastrasse 13, 49076 Osnabrück (Germany)
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  • This work was supported by the Marie Curie Actions COFUND (EMBL Interdisciplinary Postdoc (EIPOD) stipend to P.H.), the Netherlands Proteomics Center (NPC) within the Netherlands Genomics Initiative and an NPC Valorization Voucher (P.H.), the European Union (7th FP Marie-Curie ITN “SPHINGONET” to C.S., A.J.R.H., J.C.M.H.), and the DFG (TRR83 to B.B., C.S.). K.M. was a fellow of the Danish Natural Science Research Council (09-064986/FNU). We thank Y. Horibata, H. Sugimoto, and S. Cockcroft for providing plasmids, C. Versluis for high-resolution mass spectrometra, and H. Vlieg, H. Dijkstra, and T. Klimeck for technical assistance.

Abstract

original image

Bifunctional lipid technology: Cells convert externally added photoactivatable and “clickable” fatty acids into a variety of bifunctional phospholipids that can be covalently linked to their protein-binding partners by irradiation with UV light. Derivatization of the clickable group with a reporter molecule makes it possible to identify and image the lipid-bound proteins in situ (see scheme).

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