Cloning of the human thiamine transporter, a member of the folate transporter family

Binita Dutta, Wei Huang, Mariela Molero, Ramesh Kekuda, Frederick H. Leibach, Lawrence D Devoe, Vadivel Ganapathy, Puttur D Prasad

Research output: Contribution to journalArticle

137 Citations (Scopus)

Abstract

We have isolated a cDNA from human placenta, which, when expressed heterologously in mammalian cells, mediates the transport of the water- soluble vitamin thiamine. The cDNA codes for a protein of 497 amino acids containing 12 putative transmembrane domains. Northern blot analysis indicates that this transporter is widely expressed in human tissues. When expressed in HeLa cells, the cDNA induces the transport of thiamine (K(t) = 2.5 ± 0.6 μM) in a Na+-independent manner. The cDNA-mediated transport of thiamine is stimulated by an outwardly directed H+ gradient. Substrate specificity assays indicate that the transporter is specific to thiamine. Even though thiamine is an organic cation, the cDNA-induced thiamine transport is not inhibited by other organic cations. Similarly, thiamine is not a substrate for the known members of mammalian organic cation transporter family. The thiamine transporter gene, located on human chromosome 1q24, consists of 6 exons and is most likely the gene defective in the metabolic disorder, thiamine-responsive megaloblastic anemia. At the level of amino acid sequence, the thiamine transporter is most closely related to the reduced-folate transporter and thus represents the second member of the folate transporter family.

Original languageEnglish (US)
Pages (from-to)31925-31929
Number of pages5
JournalJournal of Biological Chemistry
Volume274
Issue number45
DOIs
StatePublished - Nov 5 1999

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Folic Acid Transporters
Cloning
Thiamine
Organism Cloning
Complementary DNA
Cations
Genes
Megaloblastic Anemia
Amino Acids
Human Chromosomes
Substrates
Chromosomes
Substrate Specificity
HeLa Cells
Vitamins
Northern Blotting
Placenta

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Cloning of the human thiamine transporter, a member of the folate transporter family. / Dutta, Binita; Huang, Wei; Molero, Mariela; Kekuda, Ramesh; Leibach, Frederick H.; Devoe, Lawrence D; Ganapathy, Vadivel; Prasad, Puttur D.

In: Journal of Biological Chemistry, Vol. 274, No. 45, 05.11.1999, p. 31925-31929.

Research output: Contribution to journalArticle

Dutta, Binita ; Huang, Wei ; Molero, Mariela ; Kekuda, Ramesh ; Leibach, Frederick H. ; Devoe, Lawrence D ; Ganapathy, Vadivel ; Prasad, Puttur D. / Cloning of the human thiamine transporter, a member of the folate transporter family. In: Journal of Biological Chemistry. 1999 ; Vol. 274, No. 45. pp. 31925-31929.
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