Handling of intracellular K⁺ determines voltage dependence of plasmalemmal monoamine transporter function

The concentrative power of the transporters for dopamine (DAT), norepinephrine (NET), and serotonin (SERT) is thought to be fueled by the transmembrane Na⁺ gradient, but it is conceivable that they can also tap other energy sources, for example, membrane voltage and/or the transmembrane K⁺ gradient. We have addressed this by recording uptake of endogenous substrates or the fluorescent substrate APP⁺(4-(4-dimethylamino)phenyl-1-methylpyridinium) under voltage control in cells expressing DAT, NET, or SERT. We have shown that DAT and NET differ from SERT in intracellular handling of K⁺. In DAT and NET, substrate uptake was voltage-dependent due to the transient nature of intracellular K⁺ binding, which precluded K⁺ antiport. SERT, however, antiports K⁺ and achieves voltage-independent transport. Thus, there is a trade-off between maintaining constant uptake and harvesting membrane potential for concentrative power, which we conclude to occur due to subtle differences in the kinetics of co-substrate ion binding in closely related transporters.