Na+ - and CI- -coupled active transport of carnitine by the amino acid transporter ATB0,+ from mouse colon expressed in HRPE cells and Xenopus oocytes

Takeo Nakanishi, Takahiro Hatanaka, Wei Huang, Puttur D Prasad, Frederick H. Leibach, Malliga E. Ganapathy, Vadivel Ganapathy

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Abstract

1. ATB0,+ is an amino acid transporter energized by transmembrane gradients of Na+ and Cl- and membrane potential. We cloned this transporter from mouse colon and expressed the clone functionally in mammalian (human retinal pigment epithelial, HRPE) cells and Xenopus laevis oocytes to investigate the interaction of carnitine and its acyl esters with the transporter. 2. When expressed in mammalian cells, the cloned ATB0,+ was able to transport carnitine, propionylcarnitine and acetylcarnitine. The transport process was Na+ and Cl- dependent and inhibitable by the amino acid substrates of the transporter. The Michaelis constant for carnitine was 0.83 ± 0.08 mM and the Hill coefficient for Na+ activation was 1.6 ± 0.1. 3. When expressed in Xenopus laevis oocytes, the cloned ATB0,+ was able to induce inward currents in the presence of carnitine and propionylcarnitine under voltage-clamped conditions. There was no detectable current in the presence of acetylcarnitine. Carnitine-induced currents were obligatorily dependent on the presence of Na+ and Cl-. The currents were saturable with carnitine and the Michaelis constant was 1.8 ± 0.4 mM. The analysis of Na+-and Cl-activation kinetics revealed that 2 Na+ and 1 Cl- were involved in the transport of carnitine via the transporter. 4. These studies describe the identification of a novel function for the amino acid transporter ATB0,+. Since this transporter is expressed in the intestinal tract, lung and mammary gland, it is likely to play a significant role in the handling of carnitine in these tissues. 5. A Na+-dependent transport system for carnitine has already been described. This transporter, known as OCTN2 (novel organic cation transporter 2), is expressed in most tissues and transports carnitine with high affinity. It is energized, however, only by a Na+ gradient and membrane potential. In contrast, ATB0,+ is a low-affinity transporter for carnitine, but exhibits much higher concentrative capacity than OCTN2 because of its energization by transmembrane gradients of Na+ and Cl- as well as by membrane potential.

Original languageEnglish (US)
Pages (from-to)297-304
Number of pages8
JournalJournal of Physiology
Volume532
Issue number2
DOIs
StatePublished - Apr 15 2001

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Amino Acid Transport Systems
Retinal Pigments
Carnitine
Active Biological Transport
Xenopus
Oocytes
Colon
Epithelial Cells
propionylcarnitine
Acetylcarnitine
Membrane Potentials
Xenopus laevis
Cations
Intestinal Mucosa
Human Mammary Glands
Esters
Clone Cells

ASJC Scopus subject areas

  • Physiology

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Na+ - and CI- -coupled active transport of carnitine by the amino acid transporter ATB0,+ from mouse colon expressed in HRPE cells and Xenopus oocytes. / Nakanishi, Takeo; Hatanaka, Takahiro; Huang, Wei; Prasad, Puttur D; Leibach, Frederick H.; Ganapathy, Malliga E.; Ganapathy, Vadivel.

In: Journal of Physiology, Vol. 532, No. 2, 15.04.2001, p. 297-304.

Research output: Contribution to journalArticle

Nakanishi, Takeo ; Hatanaka, Takahiro ; Huang, Wei ; Prasad, Puttur D ; Leibach, Frederick H. ; Ganapathy, Malliga E. ; Ganapathy, Vadivel. / Na+ - and CI- -coupled active transport of carnitine by the amino acid transporter ATB0,+ from mouse colon expressed in HRPE cells and Xenopus oocytes. In: Journal of Physiology. 2001 ; Vol. 532, No. 2. pp. 297-304.
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AU - Nakanishi, Takeo

AU - Hatanaka, Takahiro

AU - Huang, Wei

AU - Prasad, Puttur D

AU - Leibach, Frederick H.

AU - Ganapathy, Malliga E.

AU - Ganapathy, Vadivel

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N2 - 1. ATB0,+ is an amino acid transporter energized by transmembrane gradients of Na+ and Cl- and membrane potential. We cloned this transporter from mouse colon and expressed the clone functionally in mammalian (human retinal pigment epithelial, HRPE) cells and Xenopus laevis oocytes to investigate the interaction of carnitine and its acyl esters with the transporter. 2. When expressed in mammalian cells, the cloned ATB0,+ was able to transport carnitine, propionylcarnitine and acetylcarnitine. The transport process was Na+ and Cl- dependent and inhibitable by the amino acid substrates of the transporter. The Michaelis constant for carnitine was 0.83 ± 0.08 mM and the Hill coefficient for Na+ activation was 1.6 ± 0.1. 3. When expressed in Xenopus laevis oocytes, the cloned ATB0,+ was able to induce inward currents in the presence of carnitine and propionylcarnitine under voltage-clamped conditions. There was no detectable current in the presence of acetylcarnitine. Carnitine-induced currents were obligatorily dependent on the presence of Na+ and Cl-. The currents were saturable with carnitine and the Michaelis constant was 1.8 ± 0.4 mM. The analysis of Na+-and Cl-activation kinetics revealed that 2 Na+ and 1 Cl- were involved in the transport of carnitine via the transporter. 4. These studies describe the identification of a novel function for the amino acid transporter ATB0,+. Since this transporter is expressed in the intestinal tract, lung and mammary gland, it is likely to play a significant role in the handling of carnitine in these tissues. 5. A Na+-dependent transport system for carnitine has already been described. This transporter, known as OCTN2 (novel organic cation transporter 2), is expressed in most tissues and transports carnitine with high affinity. It is energized, however, only by a Na+ gradient and membrane potential. In contrast, ATB0,+ is a low-affinity transporter for carnitine, but exhibits much higher concentrative capacity than OCTN2 because of its energization by transmembrane gradients of Na+ and Cl- as well as by membrane potential.

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