Calmodulin kinase II interacts with the dopamine transporter C terminus to regulate amphetamine-induced reverse transport

TY - JOUR

T1 - Calmodulin kinase II interacts with the dopamine transporter C terminus to regulate amphetamine-induced reverse transport

AU - Fog, Jacob U

AU - Khoshbouei, Habibeh

AU - Holy, Marion

AU - Owens, William A

AU - Vaegter, Christian Bjerggaard

AU - Sen, Namita

AU - Nikandrova, Yelyzaveta

AU - Bowton, Erica

AU - McMahon, Douglas G

AU - Colbran, Roger J

AU - Daws, Lynette C

AU - Sitte, Harald H

AU - Javitch, Jonathan A

AU - Galli, Aurelio

AU - Gether, Ulrik

PY - 2006

Y1 - 2006

N2 - Efflux of dopamine through the dopamine transporter (DAT) is critical for the psychostimulatory properties of amphetamines, but the underlying mechanism is unclear. Here we show that Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) plays a key role in this efflux. CaMKIIalpha bound to the distal C terminus of DAT and colocalized with DAT in dopaminergic neurons. CaMKIIalpha stimulated dopamine efflux via DAT in response to amphetamine in heterologous cells and in dopaminergic neurons. CaMKIIalpha phosphorylated serines in the distal N terminus of DAT in vitro, and mutation of these serines eliminated the stimulatory effects of CaMKIIalpha. A mutation of the DAT C terminus impairing CaMKIIalpha binding also impaired amphetamine-induced dopamine efflux. An in vivo role for CaMKII was supported by chronoamperometry measurements showing reduced amphetamine-induced dopamine efflux in response to the CaMKII inhibitor KN93. Our data suggest that CaMKIIalpha binding to the DAT C terminus facilitates phosphorylation of the DAT N terminus and mediates amphetamine-induced dopamine efflux.

AB - Efflux of dopamine through the dopamine transporter (DAT) is critical for the psychostimulatory properties of amphetamines, but the underlying mechanism is unclear. Here we show that Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) plays a key role in this efflux. CaMKIIalpha bound to the distal C terminus of DAT and colocalized with DAT in dopaminergic neurons. CaMKIIalpha stimulated dopamine efflux via DAT in response to amphetamine in heterologous cells and in dopaminergic neurons. CaMKIIalpha phosphorylated serines in the distal N terminus of DAT in vitro, and mutation of these serines eliminated the stimulatory effects of CaMKIIalpha. A mutation of the DAT C terminus impairing CaMKIIalpha binding also impaired amphetamine-induced dopamine efflux. An in vivo role for CaMKII was supported by chronoamperometry measurements showing reduced amphetamine-induced dopamine efflux in response to the CaMKII inhibitor KN93. Our data suggest that CaMKIIalpha binding to the DAT C terminus facilitates phosphorylation of the DAT N terminus and mediates amphetamine-induced dopamine efflux.

KW - Amphetamines

KW - Animals

KW - Animals, Newborn

KW - Benzylamines

KW - Biological Transport

KW - Blotting, Western

KW - Calcium-Calmodulin-Dependent Protein Kinase Type 2

KW - Calcium-Calmodulin-Dependent Protein Kinases

KW - Cells, Cultured

KW - Central Nervous System Stimulants

KW - Chromatography, High Pressure Liquid

KW - Dopamine

KW - Dopamine Plasma Membrane Transport Proteins

KW - Enzyme Inhibitors

KW - Humans

KW - Immunohistochemistry

KW - Immunoprecipitation

KW - Membrane Potentials

KW - Mesencephalon

KW - Neurons

KW - Patch-Clamp Techniques

KW - Phosphorylation

KW - Protein Binding

KW - Protein Structure, Tertiary

KW - Rats

KW - Sulfonamides

KW - Transfection

U2 - 10.1016/j.neuron.2006.06.028

DO - 10.1016/j.neuron.2006.06.028

M3 - Journal article

C2 - 16908408

SN - 0896-6273

VL - 51

SP - 417

EP - 429

JO - Neuron

JF - Neuron

IS - 4

ER -