Involvement of threonine 258 and serine 259 motif in amphetamine-induced norepinephrine transporter endocytosis.

Journal of Neurochemistry
Balasubramaniam AnnamalaiLankupalle D Jayanthi

Abstract

D-amphetamine (AMPH) down-regulates the norepinephrine transporter (NET), although the exact trafficking pathways altered and motifs involved are not known. Therefore, we examined the cellular and molecular mechanisms involved in AMPH-induced NET regulation in human placental trophoblast cells expressing the wild-type (WT)-hNET and the hNET double mutant (DM)-bearing protein kinase C (PKC)-resistant T258A + S259A motif. NET function and surface expression were significantly reduced in cells expressing WT-hNET but not in cells expressing hNET-DM following AMPH treatment. AMPH inhibited plasma membrane recycling of both WT-hNET and hNET-DM. In contrast, AMPH stimulated endocytosis of WT-hNET, and did not affect hNET-DM endocytosis. Although PKC or calcium/calmodulin- dependent kinase-II (CaMKII) inhibition or depletion of calcium failed to block AMPH-mediated down-regulation of WT-hNET, NET-specific blocker desipramine completely prevented AMPH-induced down-regulation. Furthermore, AMPH treatment had no effect on phospho-CaMKII immunoreactivity. The inhibitory potency of AMPH was highest on hNET-DM, intermediary on T258A and S259A single mutants and lowest on WT-hNET. Single mutants exhibited partial resistance to AMPH-mediated d...Continue Reading

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Citations

Dec 18, 2013·Pflügers Archiv : European journal of physiology·Gary RudnickFrancois Verrey
Sep 4, 2016·Pharmacological Reviews·Daniel P Bermingham, Randy D Blakely
Aug 28, 2013·The Journal of Biological Chemistry·Obulakshmi ArapulisamyLankupalle D Jayanthi
Jul 15, 2011·Pharmacological Reviews·Anders S KristensenUlrik Gether

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