Transport mechanisms of the imino acid L-proline in the human intestinal epithelial Caco-2 cell line

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Abstract

The intestinal transport of L-proline (L-Pro) has been investigated in various animal species with the use of different tissue preparations. Because major qualitative differences have been observed among the species, it is difficult to extent the results obtained with animal models to humans. In addition, studies on human tissue are lacking because of difficulties in obtaining material for experiments. To characterize the mechanisms involved in the intestinal absorption of L-Pro in humans, the transport of this nonessential imino acid was studied in monolayers of human intestinal Caco-2 ceils that were cultivated on microporous membranes. In this model, L-Pro was transported selectively in the apical (AP)-to-basolateral (BL) direction. This transport was significantly reduced by metabolic inhibitors and by an incubation at 4°C; it was Na+ dependent and showed competition with (methyP amino)-α-isobutyric acid and L-hydroxyproline. By contrast, transport in the BL-to-AP direction resulted to a large extent from passive movement (paracellular passage and transcellular diffusion). L-Pro accumulation by Caco-2 cells was significantly greater from the AP pole than from the BL pole. About 30-50% of the accumulated molecules were incorporated into newly synthesized proteins in a process inhibited by cycloheximide, whereas the remainder were extensively metabolized into non-amino acid compounds. L-Pro accumulations from the AP and BL poles were both Na+ dependent, but they exhibited different characteristics. AP accumulation was inhibited by competition with (methylamino)-α-isobutyric acid, L-hydroxyproline and, to a lesser extent, D-Pro, whereas BL accumulation was inhibited by competition with L-hydroxyproline, (methylamino)-α-isobutyric acid, α-aminoisobutyric acid, L-histidine and small neutral amino acids. The results indicate that AP-to-BL transport and AP accumulation of L-Pro exhibited very different characteristics than BL-to-AP transport and BL accumulation.

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Berger, V., De Bremaeker, N., Larondelle, Y., Trouet, A., & Schneider, Y. J. (2000). Transport mechanisms of the imino acid L-proline in the human intestinal epithelial Caco-2 cell line. Journal of Nutrition, 130(11), 2772–2779. https://doi.org/10.1093/jn/130.11.2772

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