A search for ceramide binding proteins using bifunctional lipid analogs yields CERT-related protein StarD7

Svenja Bockelmann, John G. M. Mina, Sergei Korneev, Dina G. Hassan, Dagmar Mueller, Angelika Hilderink, Hedwich C. Vlieg, Reinout Raijmakers, Albert J. R. Heck, Per Haberkant, Joost C. M. Holthuis

    Research output: Contribution to journalArticle

    Abstract

    Ceramides are central intermediates of sphingolipid metabolism with dual roles as mediators of cellular stress signaling and mitochondrial apoptosis. How ceramides exert their cytotoxic effects is unclear and their poor solubility in water hampers a search for specific protein interaction partners. Here, we report the application of a photoactivatable and clickable ceramide analog, pacCer, to identify ceramide binding proteins and unravel the structural basis by which these proteins recognize ceramide. Besides capturing ceramide transfer protein (CERT) from a complex proteome, our approach yielded CERT-related steroidogenic acute regulatory protein D7 (StarD7) as novel ceramide binding protein. Previous work revealed that StarD7 is required for efficient mitochondrial import of phosphatidylcholine (PC) and serves a critical role in mitochondrial function and morphology. Combining site-directed mutagenesis and photoaffinity labeling experiments, we demonstrate that the steroidogenic acute regulatory transfer domain of StarD7 harbors a common binding site for PC and ceramide. While StarD7 lacks robust ceramide transfer activity in vitro, we find that its ability to shuttle PC between model membranes is specifically affected by ceramides. Besides demonstrating the suitability of pacCer as a tool to hunt for ceramide binding proteins, our data point at StarD7 as a candidate effector protein by which ceramides may exert part of their mitochondria-mediated cytotoxic effects.
    Original languageEnglish
    Pages (from-to)515-530
    Number of pages16
    JournalJournal of Lipid Research
    Volume59
    Issue number3
    DOIs
    Publication statusPublished - 1 Mar 2018

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  • Cite this

    Bockelmann, S., Mina, J. G. M., Korneev, S., Hassan, D. G., Mueller, D., Hilderink, A., Vlieg, H. C., Raijmakers, R., Heck, A. J. R., Haberkant, P., & Holthuis, J. C. M. (2018). A search for ceramide binding proteins using bifunctional lipid analogs yields CERT-related protein StarD7. Journal of Lipid Research, 59(3), 515-530. https://doi.org/10.1194/jlr.M082354