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 journalArticle

64 Citations (Scopus)

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)
JournalAmerican Journal of Physiology - Cell Physiology
Volume278
Issue number5 47-5
StatePublished - Jun 22 2000
Externally publishedYes

Fingerprint

Dicarboxylic Acid Transporters
Succinic Acid
Induced currents
Citric Acid
Stoichiometry
Oocytes
Rats
Genes
Functional analysis
Xenopus laevis
Substrates
Chromosomes
Fluorescence In Situ Hybridization
Titration
Membrane Potentials
Introns
Placenta
Charge transfer
Exons
Catalytic Domain

Keywords

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

ASJC Scopus subject areas

  • Physiology
  • Cell Biology

Cite this

Structure, function, and genomic organization of human Na+-dependent high-affinity dicarboxylate transporter. / Wang, Haiping; Fei, You Jun; Kekuda, Ramesh; Yang-Feng, Teresa L.; Devoe, Lawrence D; Leibach, Frederick H.; Prasad, Puttur D; Ganapathy, Vadivel.

In: American Journal of Physiology - Cell Physiology, Vol. 278, No. 5 47-5, 22.06.2000.

Research output: Contribution to journalArticle

Wang, Haiping ; Fei, You Jun ; Kekuda, Ramesh ; Yang-Feng, Teresa L. ; Devoe, Lawrence D ; Leibach, Frederick H. ; Prasad, Puttur D ; Ganapathy, Vadivel. / Structure, function, and genomic organization of human Na+-dependent high-affinity dicarboxylate transporter. In: American Journal of Physiology - Cell Physiology. 2000 ; Vol. 278, No. 5 47-5.
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AU - Fei, You Jun

AU - Kekuda, Ramesh

AU - Yang-Feng, Teresa L.

AU - Devoe, Lawrence D

AU - Leibach, Frederick H.

AU - Prasad, Puttur D

AU - Ganapathy, Vadivel

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KW - Electrophysiology

KW - Exon-intron organization

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