X-ray structure of dopamine transporter elucidates antidepressant mechanism

Nature. 2013 Nov 7;503(7474):85-90. doi: 10.1038/nature12533. Epub 2013 Sep 15.

Abstract

Antidepressants targeting Na(+)/Cl(-)-coupled neurotransmitter uptake define a key therapeutic strategy to treat clinical depression and neuropathic pain. However, identifying the molecular interactions that underlie the pharmacological activity of these transport inhibitors, and thus the mechanism by which the inhibitors lead to increased synaptic neurotransmitter levels, has proven elusive. Here we present the crystal structure of the Drosophila melanogaster dopamine transporter at 3.0 Å resolution bound to the tricyclic antidepressant nortriptyline. The transporter is locked in an outward-open conformation with nortriptyline wedged between transmembrane helices 1, 3, 6 and 8, blocking the transporter from binding substrate and from isomerizing to an inward-facing conformation. Although the overall structure of the dopamine transporter is similar to that of its prokaryotic relative LeuT, there are multiple distinctions, including a kink in transmembrane helix 12 halfway across the membrane bilayer, a latch-like carboxy-terminal helix that caps the cytoplasmic gate, and a cholesterol molecule wedged within a groove formed by transmembrane helices 1a, 5 and 7. Taken together, the dopamine transporter structure reveals the molecular basis for antidepressant action on sodium-coupled neurotransmitter symporters and elucidates critical elements of eukaryotic transporter structure and modulation by lipids, thus expanding our understanding of the mechanism and regulation of neurotransmitter uptake at chemical synapses.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antidepressive Agents, Tricyclic / chemistry
  • Antidepressive Agents, Tricyclic / metabolism
  • Antidepressive Agents, Tricyclic / pharmacology*
  • Bacterial Proteins / chemistry
  • Binding Sites
  • Cholesterol / chemistry
  • Cholesterol / metabolism
  • Crystallization
  • Crystallography, X-Ray
  • Dopamine Plasma Membrane Transport Proteins / chemistry*
  • Dopamine Plasma Membrane Transport Proteins / metabolism
  • Drosophila melanogaster / chemistry*
  • Ions / chemistry
  • Ions / metabolism
  • Models, Molecular
  • Nortriptyline / chemistry
  • Nortriptyline / metabolism
  • Nortriptyline / pharmacology*
  • Protein Conformation
  • Protein Stability
  • Structure-Activity Relationship
  • Temperature

Substances

  • Antidepressive Agents, Tricyclic
  • Bacterial Proteins
  • Dopamine Plasma Membrane Transport Proteins
  • Ions
  • Cholesterol
  • Nortriptyline

Associated data

  • PDB/4M48