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Am J Physiol Gastrointest Liver Physiol 284: G863-G871, 2003. First published January 10, 2003; doi:10.1152/ajpgi.00220.2002
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Vol. 284, Issue 5, G863-G871, May 2003

Expression and localization of organic cation/carnitine transporter OCTN2 in Caco-2 cells

Ihsan Elimrani1, Karim Lahjouji1, Ernest Seidman2, Marie-Josée Roy2, Grant A. Mitchell1, and Ijaz Qureshi1

Divisions of 1 Medical Genetics and 2 Gastroenterology, Research Center, Hôpital Sainte-Justine, Department of Pediatrics, Université de Montréal, Montreal, Quebec, Canada H3T 1C5

L-Carnitine is derived both from dietary sources and biosynthesis. Dietary carnitine is absorbed in the small intestine and then distributed to other organs. Previous studies using Caco-2 cells demonstrated that the transport of L-carnitine in the intestine involves a carrier-mediated system. The purpose of this study was to determine whether the uptake of L-carnitine in Caco-2 cells is mediated by the recently identified organic cation/carnitine transporter (OCTN2). Kinetics of L-[3H]carnitine uptake were investigated with or without specific inhibitors. L-Carnitine uptake in mature cells was sodium dependent and linear with time. Km and Vmax values for saturable uptake were 14.07 ± 1.70 µM and 26.3 ± 0.80 pmol · mg protein-1 · 6 min-1, respectively. L-carnitine uptake was inhibited (P < 0.05-0.01) by valproate and other organic cations. Anti-OCTN2 antibodies recognized a protein in the brush-border membrane (BBM) of Caco-2 cells with an apparent molecular mass of 60 kDa. The OCTN2 expression was confirmed by double immunostaining. Our results demonstrate that L-carnitine uptake in differentiated Caco-2 cells is primarily mediated by OCTN2, located on the BBM.

intestine; brush-border membrane; organic cation transport; anti-OCTN2 antibodies; valproate.


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