Transport mechanisms for vitamin C in the JAR human placental choriocarcinoma cell line

Puttur D Prasad, Wei Huang, Haiping Wang, Frederick H. Leibach, Vadivel Ganapathy

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

We investigated the transport pathways available for the uptake of vitamin C in the human placental choriocarcinoma cell line, JAR. These cells were found to possess the capacity to accumulate the vitamin when presented either in the oxidized form (dehydroascorbic acid) or in the reduced form (ascorbate). Dithiothreitol and 5,5'-dithiobis(2-nitrobenzoic acid) were used to maintain vitamin C as ascorbate and dehydroascorbic acid, respectively. The uptake of these two forms of vitamin C in JAR cells was found to occur by different mechanisms. The uptake of the dehydroascorbic acid was Na+- independent and was mediated by facilitative glucose transporters as evidenced from the inhibition of the uptake process by glucose. On the other hand, the uptake of ascorbate was Na+-dependent and was not sensitive to inhibition by glucose. Substitution of Na+ with other monovalent cations abolished the uptake of ascorbate completely. The uptake process was, however, not influenced by anions. Kinetic analysis indicated the presence of a single saturable transport system for ascorbate with a Michaelis-Menten constant of 22 ± 1 μM. The dependence of the uptake rate of ascorbate on Na+ concentration exhibited sigmoidal kinetics, suggesting interaction of more than one Na+ ion with the transporter. The Hill coefficient for the Na+ interaction was 2, indicating that the Na+-dependent ascorbate transport is electrogenic. The Na+-dependent stimulation of ascorbate uptake was primarily due to an increase in the affinity of the transporter for ascorbate in the presence of Na+. It is concluded that the JAR placental trophoblast cell line expresses two different transport systems for vitamin C: one for the reduced form of the vitamin ascorbate; and the other for the oxidized form of the vitamin dehydroascorbic acid.

Original languageEnglish (US)
Pages (from-to)141-151
Number of pages11
JournalBiochimica et Biophysica Acta - Biomembranes
Volume1369
Issue number1
DOIs
StatePublished - Feb 2 1998

Fingerprint

Dehydroascorbic Acid
Choriocarcinoma
Ascorbic Acid
Cells
Vitamins
Cell Line
Dithionitrobenzoic Acid
Glucose
Monovalent Cations
Kinetics
Facilitative Glucose Transport Proteins
Dithiothreitol
Trophoblasts
Anions
Substitution reactions
Ions

Keywords

  • (Human placenta)
  • Ascorbate
  • Dehydroascorbic acid
  • JAR choriocarcinoma cell
  • Vitamin transport

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Cell Biology

Cite this

Transport mechanisms for vitamin C in the JAR human placental choriocarcinoma cell line. / Prasad, Puttur D; Huang, Wei; Wang, Haiping; Leibach, Frederick H.; Ganapathy, Vadivel.

In: Biochimica et Biophysica Acta - Biomembranes, Vol. 1369, No. 1, 02.02.1998, p. 141-151.

Research output: Contribution to journalArticle

Prasad, Puttur D ; Huang, Wei ; Wang, Haiping ; Leibach, Frederick H. ; Ganapathy, Vadivel. / Transport mechanisms for vitamin C in the JAR human placental choriocarcinoma cell line. In: Biochimica et Biophysica Acta - Biomembranes. 1998 ; Vol. 1369, No. 1. pp. 141-151.
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