Zn2+ site engineering at the oligomeric interface of the dopamine transporter

FEBS Letters

Norgaard-Nielsen, Kristine¹  Hastrup, Hanne²  Javitch, Jonathan A²  Norregaard, Lene¹  Gether, Ulrik¹ 

[1]Molecular Neuropharmacology Group, Department of Pharmacology 12-5-22, The Panum Institute, University of Copenhagen, DK-2200 Copenhagen N, Denmark

[2]Center for Molecular Recognition and Department of Pharmacology, Columbia University College of Physicians and Surgeons, New York, NY, USA

关键词: Monoamine transporter;    Na+/Cl−-dependent neurotransmitter transporter;    Oligomerization;    Dimerization;    Metal ion binding site;    hDAT;    human dopamine transporter;    NET;    norepinephrine transporter;    SERT;    serotonin transporter;    WT;    wild-type;    TM;    transmembrane segment;   

DOI  :  10.1016/S0014-5793(02)03008-9

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Increasing evidence suggests that Na⁺/Cl⁻-dependent neurotransmitter transporters exist as homo-oligomeric proteins. However, the functional implication of this oligomerization remains unclear. Here we demonstrate the engineering of a Zn²⁺ binding site at the predicted dimeric interface of the dopamine transporter (DAT) corresponding to the external end of transmembrane segment 6. Upon binding to this site, which involves a histidine inserted in position 310 (V310H) and the endogenous Cys306 within the same DAT molecule, Zn²⁺ potently inhibits [³H]dopamine uptake. These data provide indirect evidence that conformational changes critical for the translocation process may occur at the interface between two transporter molecules in the oligomeric structure.

【 授权许可】

Unknown