Structure, function, and genomic organization of human Na+-dependent high-affinity dicarboxylate transporter

Haiping Wang, You Jun Fei, Ramesh Kekuda, Teresa L. Yang-Feng, Lawrence D. Devoe, Frederick H. Leibach, Puttur D. Prasad, Vadivel Ganapathy

Research output: Contribution to journalArticlepeer-review

66 Scopus citations

Abstract

We have cloned and functionally characterized the human Na+-dependent high-affinity dicarboxylate transporter (hNaDC3) from placenta. The hNaDC3 cDNA codes for a protein of 602 amino acids with 12 transmembrane domains. When expressed in mammalian cells, the cloned transporter mediates the transport of succinate in the presence of Na+ [concentration of substrate necessary for half-maximal transport (K(t)) for succinate = 20 ± 1 μM]. Dimethylsuccinate also interacts with hNaDC3. The Na+-to-succinate stoichiometry is 3:1 and concentration of Na+ necessary for half-maximal transport (K0.5(Na+)) is 49 ± 1 mM as determined by uptake studies with radiolabeled succinate. When expressed in Xenopus laevis oocytes, hNaDC3 induces Na+-dependent inward currents in the presence of succinate and dimethylsuccinate. At a membrane potential of -50 mV, K0.5(Suc) is 102 ± 20 μM and K0.5(Na+) is 22 ± 4 mM as determined by the electrophysiological approach. Simultaneous measurements of succinate-evoked charge transfer and radiolabeled succinate uptake in hNaDC3-expressing oocytes indicate a charge-to-succinate ratio of 1:1 for the transport process, suggesting a Na+-to-succinate stoichiometry of 3:1. pH titration of citrate-induced currents shows that hNaDC3 accepts preferentially the divalent anionic form of citrate as a substrate. Li+ inhibits succinate- induced currents in the presence of Na+. Functional analysis of rat-human and human-rat NaDC3 chimeric transporters indicates that the catalytic domain of the transporter lies in the carboxy-terminal half of the protein. The human NaDC3 gene is located on chromosome 20q12-13.1, as evidenced by fluorescent in situ hybridization. The gene is >80 kbp long and consists of 13 exons and 12 introns.

Original languageEnglish (US)
Pages (from-to)C1019-C1030
JournalAmerican Journal of Physiology - Cell Physiology
Volume278
Issue number5 47-5
DOIs
StatePublished - 2000

Keywords

  • Chromosomal localization
  • Electrophysiology
  • Exon-intron organization
  • Human placenta

ASJC Scopus subject areas

  • Physiology
  • Cell Biology

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