Clustered Kv2.1 decreases dopamine transporter activity and internalization

Lebowitz, Joseph J.; Pino, Jose A.; Mackie, Phillip M.; Lin, Min; Hurst, Cheyenne; Divita, Keeley; Collins, Anthony T.; Koutzoumis, Dimitri N.; Torres, Gonzalo E.; Khoshbouei, Habibeh

Abstract

The dopamine transporter (DAT) regulates dopamine neurotransmission via reuptake of dopamine released into the extracellular space. Interactions with partner proteins alter DAT function and thereby dynamically shape dopaminergic tone important for normal brain function. However, the extent and nature of these interactions are incompletely understood. Here, we describe a novel physical and functional interaction between DAT and the voltage-gated K+ channel Kv2.1 (potassium voltage-gated channel subfamily B member 1 or KCNB1). To examine the functional consequences of this interaction, we employed a combination of immunohistochemistry, immunofluorescence live-cell microscopy, co-immunoprecipitation, and electrophysiological approaches. Consistent with previous reports, we found Kv2.1 is trafficked to membrane-bound clusters observed both in vivo and in vitro in rodent dopamine neurons. Our data provide evidence that clustered Kv2.1 channels decrease DAT's lateral mobility and inhibit its internalization, while also decreasing canonical transporter activity by altering DAT's conformational equilibrium. These results suggest that Kv2.1 clusters exert a spatially discrete homeostatic braking mechanism on DAT by inducing a relative increase in inward-facing transporters. Given recent reports of Kv2.1 dysregulation in neurological disorders, it is possible that alterations in the functional interaction between DAT and Kv2.1 affect dopamine neuron activity.

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Título según WOS: ID WOS:000467394700029 Not found in local WOS DB
Título de la Revista: JOURNAL OF BIOLOGICAL CHEMISTRY
Volumen: 294
Número: 17
Editorial: AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
Fecha de publicación: 2019
Página de inicio: 6957
Página final: 6971
DOI:

10.1074/jbc.RA119.007441

Notas: ISI