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X-ray structures of Drosophila dopamine transporter in complex with nisoxetine and reboxetine

Abstract

Most antidepressants elicit their therapeutic benefits through selective blockade of Na+/Cl-coupled neurotransmitter transporters. Here we report X-ray structures of the Drosophila melanogaster dopamine transporter in complexes with the polycyclic antidepressants nisoxetine or reboxetine. The inhibitors stabilize the transporter in an outward-open conformation by occupying the substrate-binding site. These structures explain how interactions between the binding pocket and substituents on the aromatic rings of antidepressants modulate drug-transporter selectivity.

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Figure 1: Critical interactions for antidepressant recognition in the binding pocket of dDAT.
Figure 2: Comparison of ligand orientations in antidepressant-bound dDAT structures and predicted interactions in hNET and hDAT.

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Acknowledgements

We thank J. Coleman and other members of the Gouaux laboratory for helpful discussions, L. Vaskalis for assistance with figures and H. Owen for help with manuscript preparation. We acknowledge the staff of the Northeastern Collaborative Access Team at the Advanced Photon Source for assistance with data collection. This work was supported by a US National Institutes of Health Mental Health (NIH-MH) R. Kirschstein postdoctoral fellowship and a Brain and Behavior Research Foundation Young Investigator research award (K.H.W.); by a postdoctoral fellowship from the American Heart Association (A.P.); by the NIH-MH (E.G.); and by the Methamphetamine Abuse Research Center of the Oregon Health & Science University (P50DA018165 to E.G.). E.G. is supported as an investigator with the Howard Hughes Medical Institute.

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A.P., K.H.W. and E.G. designed the project. A.P. and K.H.W. performed protein purification, crystallography and biochemical assays. A.P., K.H.W. and E.G. wrote the manuscript.

Corresponding author

Correspondence to Eric Gouaux.

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The authors declare no competing financial interests.

Integrated supplementary information

Supplementary Figure 1 FoFc densities for ligands complexed with dDAT.

a, Nisoxetine (2.2 σ); b, reboxetine (2.4 s); Cyan sticks represent (S)-stereoisomer whereas magenta sticks are (R)-stereoisomer for both the drugs.

Supplementary information

Supplementary Text and Figures

Supplementary Figure 1 and Supplementary Tables 1 and 2 (PDF 310 kb)

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Penmatsa, A., Wang, K. & Gouaux, E. X-ray structures of Drosophila dopamine transporter in complex with nisoxetine and reboxetine. Nat Struct Mol Biol 22, 506–508 (2015). https://doi.org/10.1038/nsmb.3029

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