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Originally published In Press as doi:10.1074/jbc.M407913200 on September 23, 2004

J. Biol. Chem., Vol. 279, Issue 48, 50042-50049, November 26, 2004
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Identification and Characterization of a Novel Monoamine Transporter in the Human Brain*

Karen Engel, Mingyan Zhou, and Joanne Wang{ddagger}

From the Department of Pharmaceutics, University of Washington, Seattle, Washington 98195

Precise control of monoamine neurotransmitter levels in the extracellular fluids of the brain is critical in maintaining efficient and robust neurotransmission. High affinity transporters in the solute carrier SLC6A family function in removing monoamines from the neurosynaptic cleft. Emerging evidence suggests that these transporters are only one part of a system of transporters that work in concert to maintain brain homeostasis of monoamines. Here we report the cloning and characterization of a new human plasma membrane monoamine transporter, PMAT. The PMAT cDNA encodes a protein of 530 amino acid residues with 10–12 transmembrane segments. PMAT is not homologous to known neurotransmitter transporters but exhibits low homology to members of the equilibrative nucleoside transporter family. When expressed in Madin-Darby canine kidney cells and Xenopus laevis oocytes, PMAT efficiently transports serotonin (Km = 114 µM), dopamine (Km = 329 µM), and the neurotoxin 1-methyl-4-phenylpyridinium (Km = 33 µM). In contrast, there is no significant interaction of PMAT with nucleosides or nucleobases. PMAT-mediated monoamine transport does not require Na+ or Cl– but appears to be sensitive to changes in membrane potential. Northern blot analysis showed that PMAT is predominantly expressed in the human brain and widely distributed in the central nervous system. These studies demonstrate that PMAT may be a novel low affinity transporter for biogenic amines, which, under certain conditions, might supplement the role of the high affinity transporters in the brain.

Received for publication, July 14, 2004 , and in revised form, August 31, 2004.

* This work was supported by National Institutes of Health Grant GM66233. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EBI Data Bank with accession number(s) AY485959.

{ddagger} To whom correspondence should be addressed: Dept. of Pharmaceutics, University of Washington, H272J, Health Sciences Bldg., Seattle, WA 98195-7610. Tel.: 206-221-6561; Fax: 206-543-3204; E-mail: jowang{at}u.washington.edu.


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