Article abstract
Nature Neuroscience 11, 780 - 789 (2008)
Published online: 22 June 2008 | doi:10.1038/nn.2146
The binding sites for cocaine and dopamine in the dopamine transporter overlap
Thijs Beuming1, Julie Kniazeff2, Marianne L Bergmann2, Lei Shi1,3, Luis Gracia1, Klaudia Raniszewska2, Amy Hauck Newman4, Jonathan A Javitch5, Harel Weinstein1,3, Ulrik Gether2 & Claus J Loland2
Abstract
Cocaine is a widely abused substance with psychostimulant effects that are attributed to inhibition of the dopamine transporter (DAT). We present molecular models for DAT binding of cocaine and cocaine analogs constructed from the high-resolution structure of the bacterial transporter homolog LeuT. Our models suggest that the binding site for cocaine and cocaine analogs is deeply buried between transmembrane segments 1, 3, 6 and 8, and overlaps with the binding sites for the substrates dopamine and amphetamine, as well as for benztropine-like DAT inhibitors. We validated our models by detailed mutagenesis and by trapping the radiolabeled cocaine analog [3H]CFT in the transporter, either by cross-linking engineered cysteines or with an engineered Zn2+-binding site that was situated extracellularly to the predicted common binding pocket. Our data demonstrate the molecular basis for the competitive inhibition of dopamine transport by cocaine.
- Department of Physiology and Biophysics, Weill Medical College of Cornell University, New York, New York 10021, USA.
- Molecular Neuropharmacology Group, Department of Neuroscience and Pharmacology, The Faculty of Health Sciences, The Panum Institute 18.6, University of Copenhagen, Blegdamsvej 3, DK-2200 Copenhagen, Denmark.
- Institute for Computational Biomedicine, Weill Medical College of Cornell University, Room E-509, 1300 York Ave., New York, New York 10021, USA.
- Medicinal Chemistry Section, Medications Discovery Research Branch, National Institute on Drug Abuse, Intramural Research Program, Department of Health and Human Services, National Institutes of Health, 5500 Nathan Shock Dr., Baltimore, Maryland 21224, USA.
- Center for Molecular Recognition and Departments of Psychiatry and Pharmacology, College of Physicians and Surgeons, Columbia University, P&S 11-401, 630 West 168th St., New York, New York 10032, USA.
Correspondence to: Ulrik Gether2 e-mail: gether@sund.ku.dk
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