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Optical control of sphingosine-1-phosphate formation and function

Nature Chemical Biology volume 15, pages 623–631 (2019)Cite this article

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Abstract

Sphingosine-1-phosphate (S1P) plays important roles as a signaling lipid in a variety of physiological and pathophysiological processes. S1P signals via a family of G-protein-coupled receptors (GPCRs) (S1P1–5) and intracellular targets. Here, we report on photoswitchable analogs of S1P and its precursor sphingosine, respectively termed PhotoS1P and PhotoSph. PhotoS1P enables optical control of S1P1–3, shown through electrophysiology and Ca2+ mobilization assays. We evaluated PhotoS1P in vivo, where it reversibly controlled S1P3-dependent pain hypersensitivity in mice. The hypersensitivity induced by PhotoS1P is comparable to that induced by S1P. PhotoS1P is uniquely suited for the study of S1P biology in cultured cells and in vivo because it exhibits prolonged metabolic stability compared to the rapidly metabolized S1P. Using lipid mass spectrometry analysis, we constructed a metabolic map of PhotoS1P and PhotoSph. The formation of these photoswitchable lipids was found to be light dependent, providing a novel approach to optically probe sphingolipid biology.

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Fig. 1: Design, synthesis and photophysical properties of PhotoSph and PhotoS1P.
Fig. 2: Optical control of S1P1-GIRK coupling.
Fig. 3: Optical control of S1P1–5 receptor-mediated calcium release and homology modeling of S1P1–2 receptors.
Fig. 4: Optical control of nociception in DRG neurons and mice.
Fig. 5: In vitro SPHK assay.
Fig. 6: Lipid mass spectrometry analysis of PhotoSph/PhotoS1P metabolism.

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The authors declare that all relevant data supporting the findings in this study are available within this paper and the Supplementary Information files.

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Acknowledgements

J.M. thanks the German Academic Scholarship Foundation (Studienstiftung) for a PhD Fellowship. J.M. and A.J.E.N. thank New York University for MacCracken PhD fellowships. T.H. was supported by the Japan Society for the Promotion of Science Postdoctoral Fellowships for Research Abroad. D.D.N. and G.Y.T. were supported by NCI grant No. CA092160. H.R. was supported by Sinergia, the Swiss National Science Foundation (No. CRSII3-154405) and the NCCR Chemical Biology funded by the Swiss National Science Foundation (No. 51NF40-160589). D.M.B was supported by NIH grants Nos. AR059385 and NS077224, and by an HHMI Faculty Scholar Award. E.Y.I. was supported by NIH grant 1U01MH109069-01. We thank B. Hetzler for critical discussion of the photophysical characterization and C. Lin for assistance with NMR experiments. S. Lee is acknowledged for performing TNA-α assays with PhotoS1P on S1P receptor subtypes (data not included).

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Authors and Affiliations

  1. Department of Chemistry, New York University, New York, New York, NY, USA

    Johannes Morstein, Alexander J. E. Novak & Dirk Trauner

  2. Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA, USA

    Rose Z. Hill, Prashant C. Donthamsetti, Ehud Y. Isacoff & Diana M. Bautista

  3. Department of Biochemistry, University of Geneva, Geneva, Switzerland

    Suihan Feng, Takeshi Harayama & Howard Riezman

  4. National Centre of Competence in Research in Chemical Biology, University of Geneva, Geneva, Switzerland

    Suihan Feng, Takeshi Harayama & Howard Riezman

  5. Department of Physiology, College of Medicine, University of Tennessee Health Science Center, Memphis, TN, USA

    Derek D. Norman & Gabor J. Tigyi

  6. Department of Chemistry and Center for Integrated Protein Science, Ludwig Maximilians University Munich, Munich, Germany

    James A. Frank & Benjamin M. Williams

  7. Department of Chemistry, University of Memphis, Memphis, TN, USA

    Abby L. Parrill

  8. Computational Research on Materials Institute, University of Memphis, Memphis, TN, USA

    Abby L. Parrill

  9. Helen Wills Neuroscience Institute, University of California, Berkeley, Berkeley, CA, USA

    Ehud Y. Isacoff & Diana M. Bautista

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Contributions

J.M. and D.T. designed and coordinated the study with critical input from all authors. A.J.E.N. performed chemical synthesis with input from B.M.W. P.C.D. and J.M. performed electrophysiology experiments under supervision by E.Y.I. and with input from J.A.F. D.D.N. and J.M. performed Ca2+ mobilization experiments under supervision by G.J.T. A.L.P. performed receptor homology modeling and docking experiments. R.Z.H. performed DRG neuronal Ca2+ imaging and in vivo pain physiology experiments under supervision by D.B. J.M. performed SphK assays. S.F. performed lipid mass spectrometry analysis. T.H. performed whole-lipidome analysis. T.H. generated CRISPR KO cell lines under supervision by H.R. J.M. and D.T. wrote the manuscript with critical input from all authors.

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Correspondence to Dirk Trauner.

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Supplementary Tables 1 and 2, Supplementary Figs. 1–12

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Morstein, J., Hill, R.Z., Novak, A.J.E. et al. Optical control of sphingosine-1-phosphate formation and function. Nat Chem Biol 15, 623–631 (2019). https://doi.org/10.1038/s41589-019-0269-7

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