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

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.

[1]  J. Glowinski,et al.  Inhibition of Uptake of Tritiated-noradrenaline in the Intact Rat Brain by Imipramine and Structurally Related Compounds , 1964, Nature.

[2]  H. Rowe,et al.  The effect of nisoxetine (Lilly compound 94939), a potential antidepressant, on biogenic amine uptake in man. , 1976, British journal of clinical pharmacology.

[3]  E. Richelson,et al.  Blockade by antidepressants and related compounds of biogenic amine uptake into rat brain synaptosomes: most antidepressants selectively block norepinephrine uptake. , 1984, European journal of pharmacology.

[4]  D. Gehlert,et al.  Comparison of (R)‐[3H] Tomoxetine and (R/S)‐[3H] Nisoxetine Binding in Rat Brain , 1995, Journal of neurochemistry.

[5]  Z. Otwinowski,et al.  [20] Processing of X-ray diffraction data collected in oscillation mode. , 1997, Methods in enzymology.

[6]  S. Stahl,et al.  Basic psychopharmacology of antidepressants, part 1: Antidepressants have seven distinct mechanisms of action. , 1998, The Journal of clinical psychiatry.

[7]  G. Rudnick,et al.  Ion-coupled neurotransmitter transport: thermodynamic vs. kinetic determinations of stoichiometry. , 1998, Methods in enzymology.

[8]  Susan G. Amara,et al.  Reboxetine: a pharmacologically potent, selective, and specific norepinephrine reuptake inhibitor , 2000, Biological Psychiatry.

[9]  J. Fleishaker,et al.  Clinical Pharmacokinetics of Reboxetine, a Selective Norepinephrine Reuptake Inhibitor for the Treatment of Patients with Depression , 2000, Clinical pharmacokinetics.

[10]  J. Hirsh,et al.  The antidepressant-sensitive dopamine transporter in Drosophila melanogaster: a primordial carrier for catecholamines. , 2001, Molecular pharmacology.

[11]  Roland Contreras,et al.  Structure and function in rhodopsin: High-level expression of rhodopsin with restricted and homogeneous N-glycosylation by a tetracycline-inducible N-acetylglucosaminyltransferase I-negative HEK293S stable mammalian cell line , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[12]  R. Gainetdinov,et al.  Plasma membrane monoamine transporters: structure, regulation and function , 2003, Nature Reviews Neuroscience.

[13]  Manuel C. Peitsch,et al.  SWISS-MODEL: an automated protein homology-modeling server , 2003, Nucleic Acids Res..

[14]  Kevin Cowtan,et al.  research papers Acta Crystallographica Section D Biological , 2005 .

[15]  Jia Zhou.  Norepinephrine transporter inhibitors and their therapeutic potential. , 2004, Drugs of the future.

[16]  L. Iversen,et al.  Neurotransmitter transporters and their impact on the development of psychopharmacology , 2006, British journal of pharmacology.

[17]  J. Javitch,et al.  Monitoring the function of membrane transport proteins in detergent-solubilized form , 2007, Proceedings of the National Academy of Sciences.

[18]  Randy J. Read,et al.  Phaser crystallographic software , 2007, Journal of applied crystallography.

[19]  Eric Gouaux,et al.  Antidepressant binding site in a bacterial homologue of neurotransmitter transporters , 2007, Nature.

[20]  N. Grishin,et al.  PROMALS3D: a tool for multiple protein sequence and structure alignments , 2008, Nucleic acids research.

[21]  B. Ploeger,et al.  Estimation of binding rate constants using a simultaneous mixed‐effects method: application to monoamine transporter reuptake inhibitor reboxetine , 2010, British journal of pharmacology.

[22]  Harini Krishnamurthy,et al.  Neurotransmitter/sodium symporter orthologue LeuT has a single high–affinity substrate site , 2010, Nature.

[23]  U. Gether,et al.  SLC6 Neurotransmitter Transporters: Structure, Function, and Regulation , 2011, Pharmacological Reviews.

[24]  A. Sandelin,et al.  Interaction of Antidepressants with the Serotonin and Norepinephrine Transporters , 2012, The Journal of Biological Chemistry.

[25]  P. Zwart,et al.  Towards automated crystallographic structure refinement with phenix.refine , 2012, Acta crystallographica. Section D, Biological crystallography.

[26]  Eric Gouaux,et al.  X-ray structure of dopamine transporter elucidates antidepressant mechanism , 2013, Nature.

[27]  E. Gouaux,et al.  Structural basis for action by diverse antidepressants on biogenic amine transporters , 2013, Nature.

[28]  E. Gouaux,et al.  Screening and large-scale expression of membrane proteins in mammalian cells for structural studies , 2014, Nature Protocols.

[29]  E. Gouaux,et al.  Neurotransmitter and psychostimulant recognition by the dopamine transporter , 2015, Nature.