Letter | Published:

Structural insights into adiponectin receptors suggest ceramidase activity

Nature volume 544, pages 120123 (06 April 2017) | Download Citation

Abstract

Adiponectin receptors (ADIPORs) are integral membrane proteins that control glucose and lipid metabolism by mediating, at least in part, a cellular ceramidase activity1 that catalyses the hydrolysis of ceramide to produce sphingosine and a free fatty acid (FFA). The crystal structures of the two receptor subtypes, ADIPOR1 and ADIPOR2, show a similar overall seven-transmembrane-domain architecture with large unoccupied cavities and a zinc binding site within the seven transmembrane domain2. However, the molecular mechanisms by which ADIPORs function are not known. Here we describe the crystal structure of ADIPOR2 bound to a FFA molecule and show that ADIPOR2 possesses intrinsic basal ceramidase activity that is enhanced by adiponectin. We also identify a ceramide binding pose and propose a possible mechanism for the hydrolytic activity of ADIPOR2 using computational approaches. In molecular dynamics simulations, the side chains of residues coordinating the zinc rearrange quickly to promote the nucleophilic attack of a zinc-bound hydroxide ion onto the ceramide amide carbonyl. Furthermore, we present a revised ADIPOR1 crystal structure exhibiting a seven-transmembrane-domain architecture that is clearly distinct from that of ADIPOR2. In this structure, no FFA is observed and the ceramide binding pocket and putative zinc catalytic site are exposed to the inner membrane leaflet. ADIPOR1 also possesses intrinsic ceramidase activity, so we suspect that the two distinct structures may represent key steps in the enzymatic activity of ADIPORs. The ceramidase activity is low, however, and further studies will be required to characterize fully the enzymatic parameters and substrate specificity of ADIPORs. These insights into ADIPOR function will enable the structure-based design of potent modulators of these clinically relevant enzymes.

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Acknowledgements

We thank C. Mueller-Dieckmann and U. Zander at the European Synchrotron Radiation Facility (ESRF) for assistance in using beamline ID30B. We acknowledge the ESRF for provision of synchrotron radiation facilities via SSX Block Allocation Group beamtime. We thank R. Joosten and A. Perrakis from the PDB REDO server for help with ADIPOR1 data re-analysis and F. Rey from the Structural Virology Unit, Institut Pasteur for S2 cells and an expression vector for scFv. This project received funding from the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation programme (grant agreement 647687).

Author information

Author notes

    • Ieva Vasiliauskaité-Brooks
    •  & Remy Sounier

    These authors contributed equally to this work.

Affiliations

  1. Institut de Génomique Fonctionnelle, CNRS UMR-5203 INSERM U1191, University of Montpellier, 34094 Montpellier, France

    • Ieva Vasiliauskaité-Brooks
    • , Remy Sounier
    • , Pascal Rochaix
    • , Gaëtan Bellot
    • , Mathieu Fortier
    • , Cédric Leyrat
    •  & Sébastien Granier
  2. Centre de Biochimie Structurale, CNRS UMR 5048-INSERM 1054 University of Montpellier, 29 rue de Navacelles, 34090 Montpellier Cedex, France

    • François Hoh
  3. Division of Structural Biology, University of Oxford, Oxford, UK

    • Luigi De Colibus
  4. Dynamique des Interactions Membranaires Normales et Pathologiques, CNRS UMR5235, University of Montpellier, 34095 Montpellier, France

    • Chérine Bechara
  5. Institute for chemistry, Humboldt-Universität zu Berlin, Brook-Taylor-Str. 2, 12489 Berlin, Germany

    • Essa M. Saied
    •  & Christoph Arenz
  6. Chemistry Department, Faculty of Science, Suez Canal University, 41522 Ismailia, Egypt

    • Essa M. Saied

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Contributions

I.V.-B. and R.S. expressed, purified, characterized and crystallized receptor and scFv preparations with the help of P.R., G.B., M.F. and C.B. I.V.-B. and C.L. collected data with the help of F.H. C.L. performed the computational studies with help from L.D.C. I.V.-B. and C.L. solved and refined the structures. R.S. prepared the figures with the help of I.V.-B. and C.L. E.M.S. synthesized ceramides of different chain lengths and performed UPLC–MS analysis of the ceramide cleavage reactions. C.A. supervised E.M.S. All authors contributed to the manuscript preparation. S.G. supervised the project.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Cédric Leyrat or Sébastien Granier.

Reviewer Information Nature thanks D. Veprintsev and the other anonymous reviewer(s) for their contribution to the peer review of this work.

Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

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DOI

https://doi.org/10.1038/nature21714

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