1. Biochemistry and Chemical Biology
Download icon

Optical manipulation of sphingolipid biosynthesis using photoswitchable ceramides

  1. Matthijs Kol
  2. Ben Williams
  3. Henry Toombs-Ruane
  4. Henri G Franquelim
  5. Sergei Korneev
  6. Christian Schroeer
  7. Petra Schwille
  8. Dirk Trauner  Is a corresponding author
  9. Joost CM Holthuis  Is a corresponding author
  10. James A Frank  Is a corresponding author
  1. University of Osnabrück, Germany
  2. Ludwig Maximilians University Munich, Germany
  3. Max Planck Institute of Biochemistry, Germany
  4. New York University, United States
  5. University of Osnabrück, Germany
  6. Oregon Health and Science University, United States
Tools and Resources
  • Cited 1
  • Views 2,154
  • Annotations
Cite this article as: eLife 2019;8:e43230 doi: 10.7554/eLife.43230

Abstract

Ceramides are central intermediates of sphingolipid metabolism that also function as potent messengers in stress signaling and apoptosis. Progress in understanding how ceramides execute their biological roles is hampered by a lack of methods to manipulate their cellular levels and metabolic fate with appropriate spatiotemporal precision. Here, we report on clickable, azobenzene-containing ceramides, caCers, as photoswitchable metabolic substrates to exert optical control over sphingolipid production in cells. Combining atomic force microscopy on model bilayers with metabolic tracing studies in cells, we demonstrate that light-induced alterations in the lateral packing of caCers lead to marked differences in their metabolic conversion by sphingomyelin synthase and glucosylceramide synthase. These changes in metabolic rates are instant and reversible over several cycles of photoswitching. Our findings disclose new opportunities to probe the causal roles of ceramides and their metabolic derivatives in a wide array of sphingolipid-dependent cellular processes with the spatiotemporal precision of light.

Article and author information

Author details

  1. Matthijs Kol

    Department of Biology/Chemistry, University of Osnabrück, Osnabrück, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-3068-6501
  2. Ben Williams

    Department of Chemistry, Ludwig Maximilians University Munich, Munich, Germany
    Competing interests
    The authors declare that no competing interests exist.
  3. Henry Toombs-Ruane

    Department of Chemistry, Ludwig Maximilians University Munich, Munich, Germany
    Competing interests
    The authors declare that no competing interests exist.
  4. Henri G Franquelim

    Department of Cellular and Molecular Biophysics, Max Planck Institute of Biochemistry, Martinsried, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6229-4276
  5. Sergei Korneev

    Department of Biology/Chemistry, University of Osnabrück, Osnabrück, Germany
    Competing interests
    The authors declare that no competing interests exist.
  6. Christian Schroeer

    Department of Biology/Chemistry, University of Osnabrück, Osnabrück, Germany
    Competing interests
    The authors declare that no competing interests exist.
  7. Petra Schwille

    Department of Cellular and Molecular Biophysics, Max Planck Institute of Biochemistry, Martinsried, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-6106-4847
  8. Dirk Trauner

    Department of Chemistry, New York University, New York, United States
    For correspondence
    dirktrauner@nyu.edu
    Competing interests
    The authors declare that no competing interests exist.
  9. Joost CM Holthuis

    Department of Biology/Chemistry, University of Osnabrück, Osnabrück, Germany
    For correspondence
    Joost.Holthuis@Biologie.Uni-Osnabrueck.DE
    Competing interests
    The authors declare that no competing interests exist.
  10. James A Frank

    The Vollum Institute, Oregon Health and Science University, Portland, United States
    For correspondence
    frankja@ohsu.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6705-2540

Funding

Deutsche Forschungsgemeinschaft (SFB1032)

  • Henri G Franquelim
  • Petra Schwille
  • Dirk Trauner
  • James A Frank

National Sciences and Engineering Research Council of Canada

  • Ben Williams

Deutsche Forschungsgemeinschaft (SFB944)

  • Matthijs Kol
  • Joost CM Holthuis

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Arun Radhakrishnan, University of Texas Southwestern Medical Center, United States

Publication history

  1. Received: October 30, 2018
  2. Accepted: February 2, 2019
  3. Accepted Manuscript published: February 5, 2019 (version 1)
  4. Version of Record published: February 22, 2019 (version 2)

Copyright

© 2019, Kol et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 2,154
    Page views
  • 379
    Downloads
  • 1
    Citations

Article citation count generated by polling the highest count across the following sources: Crossref, PubMed Central, Scopus.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Download citations (links to download the citations from this article in formats compatible with various reference manager tools)

Open citations (links to open the citations from this article in various online reference manager services)

Further reading

    1. Biochemistry and Chemical Biology
    Sandy Mattijssen et al.
    Research Advance Updated

    La-related protein 4 (LARP4) directly binds both poly(A) and poly(A)-binding protein (PABP). LARP4 was shown to promote poly(A) tail (PAT) lengthening and stabilization of individual mRNAs presumably by protection from deadenylation (Mattijssen et al., 2017). We developed a nucleotide resolution transcriptome-wide, single molecule SM-PAT-seq method. This revealed LARP4 effects on a wide range of PAT lengths for human mRNAs and mouse mRNAs from LARP4 knockout (KO) and control cells. LARP4 effects are clear on long PAT mRNAs but become more prominent at 30–75 nucleotides. We also analyzed time courses of PAT decay transcriptome-wide and for ~200 immune response mRNAs. This demonstrated accelerated deadenylation in KO cells on PATs < 75 nucleotides and phasing consistent with greater PABP dissociation in the absence of LARP4. Thus, LARP4 shapes PAT profiles throughout mRNA lifespan with impact on mRNA decay at short lengths known to sensitize PABP dissociation in response to deadenylation machinery.

    1. Biochemistry and Chemical Biology
    2. Microbiology and Infectious Disease
    Ali Alghamdi et al.
    Research Article

    Mutations in the Trypanosoma brucei aquaporin AQP2 are associated with resistance to pentamidine and melarsoprol. We show that TbAQP2 but not TbAQP3 was positively selected for increased pore size from a common ancestor aquaporin. We demonstrate that TbAQP2's unique architecture permits pentamidine permeation through its central pore and show how specific mutations in highly conserved motifs affect drug permeation. Introduction of key TbAQP2 amino acids into TbAQP3 renders the latter permeable to pentamidine. Molecular dynamics demonstrates that permeation by dicationic pentamidine is energetically favourable in TbAQP2, driven by the membrane potential, although aquaporins are normally strictly impermeable for ionic species. We also identify the structural determinants that make pentamidine a permeant although most other diamidine drugs are excluded. Our results have wide-ranging implications for optimising antitrypanosomal drugs and averting cross-resistance. Moreover, these new insights in aquaporin permeation may allow the pharmacological exploitation of other members of this ubiquitous gene family.