biostudies-literaturePedersen CNAarhus Universitets ForskningsfondNIDA NIH HHSDanish Ministry for Research and Higher EducationNIMH NIH HHSNCI NIH HHSNational Institutes of HealthNovo Nordisk FondenLundbeck FoundationNIH HHS2043-2055https://www.ebi.ac.uk/biostudies/studies/S-EPMC11449642biostudies-literatureUnknown168(9)The regulation of dopamine (DA) removal from the synaptic cleft is a crucial process in neurotransmission and is facilitated by the sodium- and chloride-coupled dopamine transporter DAT. Psychostimulant drugs, cocaine, and amphetamine, both block the uptake of DA, while amphetamine also triggers the release of DA. As a result, they prolong or even amplify neurotransmitter signaling. Atypical inhibitors of DAT lack cocaine-like rewarding effects and offer a promising strategy for the treatment of drug use disorders. Here, we present the 3.2 Å resolution cryo-electron microscopy structure of the Drosophila melanogaster dopamine transporter (dDAT) in complex with the atypical non-competitive inhibitor AC-4-248. The inhibitor partially binds at the central binding site, extending into the extrJournal of neurochemistryCryo-EM structure of the dopamine transporter with a novel atypical non-competitive inhibitor bound to the orthosteric site.PMC11449642P30 CA010815-53R01 DA051205AUFF‐E‐2022‐9‐24NNF19OC0054875R310‐2018‐3713R01 DA051205-03P30 CA010815S10OD030245-01R346-2020-1944MH106912R01 DA051205‐03S10 OD030245MH1214535072-00025BNNF18OC0032608R01 MH106912R56 MH121453P30 CA010815‐53S10OD030245‐01R310-2018-3713NNF18OC0054875AUFF-E-2022-9-24R368-2021-522R368‐2021‐522Pedersen CNNeumann CMTDesdorf LMAkunuri RMortensen OVSchiott BYang FIta STrampari SSalvino JMShahsavar AXu YNissen PfalseCryo-EM structure of the dopamine transporter with a novel atypical non-competitive inhibitor bound to the orthosteric site.The regulation of dopamine (DA) removal from the synaptic cleft is a crucial process in neurotransmission and is facilitated by the sodium- and chloride-coupled dopamine transporter DAT. Psychostimulant drugs, cocaine, and amphetamine, both block the uptake of DA, while amphetamine also triggers the release of DA. As a result, they prolong or even amplify neurotransmitter signaling. Atypical inhibitors of DAT lack cocaine-like rewarding effects and offer a promising strategy for the treatment of drug use disorders. Here, we present the 3.2 Å resolution cryo-electron microscopy structure of the Drosophila melanogaster dopamine transporter (dDAT) in complex with the atypical non-competitive inhibitor AC-4-248. The inhibitor partially binds at the central binding site, extending into the extr2024-01-01T00:00:00Z2024 Sep2026-05-29T11:34:19.236Z2026-04-08T04:30:13.681ZS-EPMC114496423901068110.1111/jnc.16179