The dopamine transporter antiports potassium to increase the uptake of dopamine

The dopamine transporter facilitates dopamine reuptake from the extracellular space to terminate neurotransmission. The transporter belongs to the neurotransmitter:sodium symporter family, which includes transporters for serotonin, norepinephrine, and GABA that utilize the Na(+) gradient to drive the uptake of substrate. Decades ago, it was shown that the serotonin transporter also antiports K(+), but investigations of K(+)-coupled transport in other neurotransmitter:sodium symporters have been inconclusive. Here, we show that ligand binding to the Drosophila- and human dopamine transporters are inhibited by K(+), and the conformational dynamics of the Drosophila dopamine transporter in K(+) are divergent from the apo- and Na(+)-states. Furthermore, we find that K(+) increases dopamine uptake by the Drosophila dopamine transporter in liposomes, and visualize Na(+) and K(+) fluxes in single proteoliposomes using fluorescent ion indicators. Our results expand on the fundamentals of dopamine transport and prompt a reevaluation of the impact of K(+) on other transporters in this pharmacologically important family.