Transport and inhibition of the sphingosine-1-phosphate exporter SPNS2
Sphingosine-1-phosphate (S1P) is a signaling lysolipid critical to heart development, immunity, and hearing. Accordingly, mutations in the S1P transporter SPNS2 are associated with reduced white cell count and hearing defects. SPNS2 also exports the S1P-mimicking FTY720-P (Fingolimod) and thereby is central to the pharmacokinetics of this drug when treating multiple sclerosis. Here, we use a combination of cryo-electron microscopy, immunofluorescence, in vitro binding and in vivo S1P export assays, and molecular dynamics simulations to probe SPNS2's substrate binding and transport. These results reveal the transporter's binding mode to its native substrate S1P, the therapeutic FTY720-P, and the reported SPNS2-targeting inhibitor 33p. Further capturing an inward-facing apo state, our structures illuminate the protein's mechanism for exchange between inward-facing and outward-facing conformations. Finally, using these structural, localization, and S1P transport results, we identify how pathogenic mutations ablate the protein's export activity and thereby lead to hearing loss.
Authors
- Tabea Wiedmer (ORCID: https://orcid.org/0000-0002-0355-9315)
- A.C.W. Pike (ORCID: https://orcid.org/0000-0001-9661-2607)
- Benedikt Mathias Kessler (ORCID: https://orcid.org/0000-0002-8160-2446)
- Adán Pinto-Fernández (ORCID: https://orcid.org/0000-0003-1693-9664)
- Hannah C. Scott (ORCID: https://orcid.org/0009-0006-8019-9209)
- Huanyu Li (ORCID: https://orcid.org/0000-0002-5482-9936)
- Vera Puetter
- Gernot Wolf (ORCID: https://orcid.org/0000-0002-3943-8662)
- Josefina Stanka (ORCID: https://orcid.org/0009-0003-3173-703X)
- Frank Wunder (ORCID: https://orcid.org/0000-0002-1311-2357)
- Saša Štefanić (ORCID: https://orcid.org/0000-0001-7367-1831)
- Shubhashish Mukhopadhyay (ORCID: https://orcid.org/0000-0003-1430-5529)
- David B. Sauer (ORCID: https://orcid.org/0000-0001-9291-4640)
- Helena Batoulis
- Giulio Superti‐Furga (ORCID: https://orcid.org/0000-0002-0570-1768)
- Katharina Duerr (ORCID: https://orcid.org/0000-0002-3245-4088)
- N. Burgess-Brown (ORCID: https://orcid.org/0000-0003-0463-445X)
- Yung-Ning Chang (ORCID: https://orcid.org/0000-0002-9309-5955)
- Alexander Ehrmann
- Syma Khalid (ORCID: https://orcid.org/0000-0002-3694-5044)
- Zuzana Gelová (ORCID: https://orcid.org/0000-0003-4783-1752)
- Álvaro Inglés‐Prieto (ORCID: https://orcid.org/0000-0002-5409-8571)
- Andreea Scacioc (ORCID: https://orcid.org/0000-0002-7364-2843)
- Dheeraj Prakaash (ORCID: https://orcid.org/0000-0002-2209-9713)
- Andrea Garofoli
- G. McKinley (ORCID: https://orcid.org/0000-0001-5220-4337)
- C. Moreau (ORCID: https://orcid.org/0009-0000-1393-664X)
Institutions
- Austrian Academy of Sciences (AT)
- University of Zurich (CH)
- Nuffield Orthopaedic Centre (GB)
- University of Oxford (GB)
- CeMM Research Center for Molecular Medicine (AT)
- Nuvisan (Germany) (DE)
- Bayer (Germany) (DE)
- Medical University of Vienna (AT)
Publication Details
- Journal
- Nature Communications
- Published
- 2025-01-16
- DOI
- https://doi.org/10.1038/s41467-025-55942-7
- Citations
- 6
- Primary Topic
- Sphingolipid Metabolism and Signaling
- Type
- article
- Field-Weighted Citation Impact
- 4.84
Funders
- McGill University
- Wellcome Trust
- Diamond Light Source
- European Federation of Pharmaceutical Industries and Associations
- European Commission
- Universität Wien
- Medizinische Universität Wien
- Innovative Medicines Initiative
- Engineering and Physical Sciences Research Council