Abstract
The dopamine transporter (DAT) is crucial for regulating dopamine signalling and is the prime mediator for the rewarding and addictive effects of cocaine1. As part of the neurotransmitter sodium symporter family, DAT uses the Na+ gradient across cell membranes to transport dopamine against its chemical gradient2. The transport mechanism involves both intra- and extracellular gates that control substrate access to a central site. However, the molecular intricacies of this process and the inhibitory mechanism of cocaine have remained unclear. Here, we present the molecular structure of human DAT in complex with cocaine at a resolution of 2.66 Å. Our findings reveal that DAT adopts the expected LeuT-fold, posing in an outward-open conformation with cocaine bound at the central (S1) site. Notably, while an Na+ occupies the second Na+ site (Na2), the Na1 site seems to be vacant, with the side chain of Asn82 occupying the presumed Na+ space. This structural insight elucidates the mechanism for the cocaine inhibition of human DAT and deepens our understanding of neurotransmitter transport. By shedding light on the molecular underpinnings of how cocaine acts, our study lays a foundation for the development of targeted medications to combat addiction.
| Originalsprog | Engelsk |
|---|---|
| Tidsskrift | Nature |
| Vol/bind | 632 |
| Udgave nummer | 8025 |
| Sider (fra-til) | 678-685 |
| Antal sider | 8 |
| ISSN | 0028-0836 |
| DOI | |
| Status | Udgivet - 2024 |
Bibliografisk note
Funding Information:We thank L. Rosenquist for technical assistance and A. Nygaard and M. Gram for critical reading of the manuscript. The financial support for this work was provided by the Lundbeck Foundation (R344-2020-1020 to C.J.L. and R368-2021-522 to A.S.); the Independent Research Fund Denmark (3101-00381B to C.J.L.); the Carlsberg Foundation (CF20-0345 to C.J.L.); and the Maersk Foundation (L-2022-00324 to J.C.N. and L-2021-00122 to K.S.). I.E.K is the recipient of a H2020 Marie Sklodowska-Curie training network (Program number 860954). A.S. was supported by a Lundbeck Foundation Fellows grant (R368-2021-522). Cryo-EM data collection was performed at the Danish Cryo-EM Facility at the Core Facility for Integrated Microscopy (CFIM), University of Copenhagen supported by the Novo Nordisk Foundation (grants NNF17SA0024386 and NNF22OC0075808).
Funding Information:
Molecular graphics images were produced using Chimera v.1.8 from the Resource for Biocomputing, Visualization, and Informatics at the University of California, San Francisco (supported by NIH P41 RR-01081).
Publisher Copyright:
© The Author(s), under exclusive licence to Springer Nature Limited 2024.