Sodium-dependent high-affinity binding of carnitine to human placental brush border membranes

Arlene S. Roque, Puttur D Prasad, Jatinder J Bhatia, Frederick H. Leibach, Vadivel Ganapathy

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

The interaction of carnitine with human placental brush-border membrane vesicles was investigated. Carnitine was found to associate with the membrane vesicles in a Na+-dependent manner. The time course of this association did not exhibit an overshoot, which is typical of a Na+ gradient-driven transport process. The absolute requirement for Na+ was noticeable whether the association of carnitine with the vesicles was measured with a short time incubation or under equilibrium conditions, indicating Na+-dependent binding of carnitine to the human placental brush-border membranes. The binding was saturable and was of a high-affinity type with a dissociation constant of 1.37 ± 0.03 μM. Anions had little or no influence on the binding process. The binding process was specific for carnitine and its acyl derivatives. Betaine also competed for the binding process, but other structurally related compounds did not. Kinetic analyses revealed that Na+ increased the affinity of the binding process for carnitine and the Na+/carnitine coupling ratio for the binding process was 1. The dissociation constant for the interaction of Na+ with the binding of carnitine was 24 ± 4 mM. This constitutes the first report on the identification of Na+-dependent high-affinity carnitine binding in the plasma membrane of a mammalian cell. Studies with purified rat renal brush-border membrane vesicles demonstrated the presence of Na+ gradient-driven carnitine transport but no Na+-dependent carnitine binding in these membrane vesicles. In contrast, purified intestinal brush-border membrane vesicles possess neither Na+ gradient-driven carnitine transport nor Na+-dependent carnitine binding.

Original languageEnglish (US)
Pages (from-to)274-282
Number of pages9
JournalBiochimica et Biophysica Acta - Biomembranes
Volume1282
Issue number2
DOIs
StatePublished - Jul 25 1996

Fingerprint

Carnitine
Brushes
Microvilli
Sodium
Membranes
Association reactions
Betaine
Cell membranes
Anions
Rats

Keywords

  • Brush-border membrane
  • Carnitine
  • High-affinity binding
  • Human placenta
  • Sodium ion dependence

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Cell Biology

Cite this

Sodium-dependent high-affinity binding of carnitine to human placental brush border membranes. / Roque, Arlene S.; Prasad, Puttur D; Bhatia, Jatinder J; Leibach, Frederick H.; Ganapathy, Vadivel.

In: Biochimica et Biophysica Acta - Biomembranes, Vol. 1282, No. 2, 25.07.1996, p. 274-282.

Research output: Contribution to journalArticle

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