Impact of the reduced folate carrier on the accumulation of active thiamin metabolites in murine leukemia cells

Rongbao Zhao, Feng Gao, Yanhua Wang, George A. Diaz, Bruce D. Gelb, I. David Goldman

Research output: Contribution to journalArticle

70 Citations (Scopus)

Abstract

The thiamin transporter encoded by SLC19A2 and the reduced folate carrier (RFC1) share 49% homology at the protein level, but the thiamin transporter does not mediate transport of folates. By using murine leukemia cell lines that express no, normal, or high levels of RFC1, we demonstrate that RFC1 does not mediate thiamin influx. However, high level RFC1 expression substantially reduced accumulation of the active thiamin coenzyme, thiamin pyrophosphate (TPP). This decreased level of TPP, synthesized intracellularly from imported thiamin, resulted from RFC1-mediated efflux of TPP. This conclusion was supported by the following observations. (i) Efflux of intracellular TPP was increased in cells with high expression of RFC1. (ii) Methotrexate inhibits TPP influx. (iii) TPP competitively inhibits methotrexate influx. (iv) Loading cells, which overexpress RFC1 to high levels of methotrexate to inhibit competitively RFC1-mediated TPP efflux, augment TPP accumulation. (v) There was an inverse correlation between thiamin accumulation and RFC1 activity in cells grown at a physiological concentration of thiamin. The modulation of thiamin accumulation by RFC1 in murine leukemia cells suggests thai this carrier may play a role in thiamin homeostasis and could serve as a modifying factor in thiamin nutritional deficiency as well as when the high affinity thiamin transporter is mutated.

Original languageEnglish (US)
Pages (from-to)1114-1118
Number of pages5
JournalJournal of Biological Chemistry
Volume276
Issue number2
DOIs
StatePublished - Jan 12 2001

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Reduced Folate Carrier Protein
Thiamine
Thiamine Pyrophosphate
Metabolites
Leukemia
Methotrexate
Thiamine Deficiency
Cells
Coenzymes
Folic Acid
Malnutrition
Homeostasis

ASJC Scopus subject areas

  • Biochemistry

Cite this

Impact of the reduced folate carrier on the accumulation of active thiamin metabolites in murine leukemia cells. / Zhao, Rongbao; Gao, Feng; Wang, Yanhua; Diaz, George A.; Gelb, Bruce D.; Goldman, I. David.

In: Journal of Biological Chemistry, Vol. 276, No. 2, 12.01.2001, p. 1114-1118.

Research output: Contribution to journalArticle

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