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Research ArticleArticle

Molecular Cloning and Functional Characterization of a Polyspecific Organic Anion Transporter from Caenorhabditis elegans

Ronald L. George, Xiang Wu, Wei Huang, You-Jun Fei, Frederick H. Leibach and Vadivel Ganapathy
Journal of Pharmacology and Experimental Therapeutics November 1999, 291 (2) 596-603;
Ronald L. George
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Xiang Wu
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Wei Huang
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You-Jun Fei
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Frederick H. Leibach
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Vadivel Ganapathy
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Abstract

We have cloned a polyspecific organic anion transporter fromCaenorhabditis elegans and elucidated its functional characteristics. The C. elegans anion transporter (CeOAT1) codes for a protein of 526 amino acids containing 12 putative transmembrane domains. It exhibits significant homology at the level of amino acid sequence to the C. elegans organic cation transporter and to the mammalian organic cation and anion transporters. The function of CeOAT1 was investigated by expressing the transporter heterologously in mammalian cells. CeOAT1 transportsp-aminohippurate (PAH) in a Na+-independent manner. The transport mechanism appears to involve anion exchange because CeOAT1-mediated PAH transport is stimulated by a cell-to-medium concentration gradient of α-ketoglutarate or fumarate generated by coexpression in the cells of a mammalian Na+-coupled dicarboxylate transporter. CeOAT1 exhibits broad specificity, accepting anions such as folate, indomethacin, furosemide, probenecid, and benzylpenicillin as substrates. The Michaelis-Menten constant for the prototypical organic anion PAH is 0.43 ± 0.07 mM. This constitutes the first report of the molecular and functional identification of a polyspecific organic anion transporter in C. elegans.

Footnotes

  • Send reprint requests to: Dr. Vadivel Ganapathy, Department of Biochemistry and Molecular Biology, Medical College of Georgia, Augusta, GA. E-mail: vganapat{at}mail.mcg.edu

  • ↵1 This work was supported by National Institutes of Health Grant DA 10045.

  • Abbreviations:
    PAH
    p-aminohippurate
    CeOAT
    Caenorhabditis elegans organic anion transporter
    OAT
    organic anion transporter
    rNaDC3
    rat Na+-coupled dicarboxylate transporter 3
    OCT
    organic cation transporter
    OCTN
    novel organic cation transporter
    MPP+
    1-methyl-4-phenylpyridinium ion
    MPTP
    1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine
    TEA
    tetraethylammonium
    RT-PCR
    reverse transcription-polymerase chain reaction
    bp
    base pair(s)
    • Received May 25, 1999.
    • Accepted July 8, 1999.
  • The American Society for Pharmacology and Experimental Therapeutics
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Journal of Pharmacology and Experimental Therapeutics: 291 (2)
Journal of Pharmacology and Experimental Therapeutics
Vol. 291, Issue 2
1 Nov 1999
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Research ArticleArticle

Molecular Cloning and Functional Characterization of a Polyspecific Organic Anion Transporter from Caenorhabditis elegans

Ronald L. George, Xiang Wu, Wei Huang, You-Jun Fei, Frederick H. Leibach and Vadivel Ganapathy
Journal of Pharmacology and Experimental Therapeutics November 1, 1999, 291 (2) 596-603;

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Research ArticleArticle

Molecular Cloning and Functional Characterization of a Polyspecific Organic Anion Transporter from Caenorhabditis elegans

Ronald L. George, Xiang Wu, Wei Huang, You-Jun Fei, Frederick H. Leibach and Vadivel Ganapathy
Journal of Pharmacology and Experimental Therapeutics November 1, 1999, 291 (2) 596-603;
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