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Copper uptake and intracellular distribution in the human intestinal Caco-2 cell line

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Abstract

The apical uptake of 64CuCl2 was investigated in human differentiated intestinal Caco-2 cells grown on permeable supports. At pH 6.0 in the apical compartment, the uptake of copper was linear over the first 6 min and between 10 and 80 μM CuCl2 exhibited non-saturable transport kinetics. In addition, copper uptake was energy-independent, affected by the valency state of copper, preferring Cu(II) over Cu(I), and not influenced by high (10 mM) extracellular calcium. The intracellular distribution of copper was investigated by FPLC at different times of uptake (`pulse') and of `chase'. Intracellular copper initially bound predominantly to low molecular weight components (i.e., glutathione), and subsequently shifted to higher molecular weight components such as metallothionein and Cu,Zn superoxide dismutase.

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References

  • Alvarez-Hernandez X, Smith M, Glass J. 1998 The effect of apotransferrin on iron release from Caco-2 cells, an intestinal epithelial cell line. Blood91, 3974–3979.

    Google Scholar 

  • Andrews N, Fleming m, Gunshin H. 1999 Iron transport across biological membranes. Nutr Rev57, 114–123.

    Google Scholar 

  • Artursson P. 1990 Epithelial transport of drugs in cell culture. I. A model for studying the passive diffusion of drugs over intestinal absorptive (Caco 2) cells. J Pharm Sci79, 476–482.

    Google Scholar 

  • Culotta V, Klomp L, Strain J, Casareno R, Krems B, Gitlin J. 1997 The copper chaperone for superoxide dismutase. J Biol Chem272, 23469–23472.

    Google Scholar 

  • Duizer E, Gilde A, Versantvoort C, Groten J. 1999 Effects of cadmium chloride on the paracellular barrier function of intestinal epithelial cells. Toxicol Appl Pharmacol155, 117–126.

    Google Scholar 

  • Ferruzza S, Ranaldi G, Di Girolamo M, Sambuy Y. 1997 The efflux of lysine from the basolateral membrane of human cultured intestinal cells (Caco-2) occurs by different mechanisms depending on the extracellular availability of amino acids. J Nutrition127, 1183–1190.

    Google Scholar 

  • Ferruzza S, Sambuy Y, Rotilio G, Ciriolo M, Scarino ML. 1999a The effect of copper on tight junctional permeability in a human intestinal cell line (Caco-2). In: Leone A, Mercer J, eds. Copper Transport and Its Disorders: Molecular and Cellular Aspects, Vol. 448. New York: Plenum Press, 215–222.

    Google Scholar 

  • Ferruzza S, Scarino ML, Rotilio G et al. 1999b Copper treatment alters the permeability of tight junctions in cultured human intestinal Caco-2 cells. Am J Physiol277, G1138–G1148.

    Google Scholar 

  • Fleming M, Trenor C, Su M et al. 1997 Microcytic anaemia mice have a mutation in Nramp2, a candidate iron transporter gene. Nat Genet16, 383–386.

    Google Scholar 

  • Freedman J, Ciriolo M, Peisach J. 1989 The role of glutathione in copper metabolism and toxicity. J Biol Chem264, 5598–5605.

    Google Scholar 

  • Gunshin H, Mackenzie B, Berger U et al. 1997Cloning and characterization of a mammalian proton-coupled metal-ion transporter. Nature338, 482–488.

    Google Scholar 

  • Han O, Failla M, Hill A, Morris E, Smith J. 1995 Reduction of Fe(III) is required for uptake of non-heme iron by Caco-2 cells. J Nutr125, 1291–1299.

    Google Scholar 

  • Linder M, Hazegh-Azam M. 1996 Copper biochemisty and molecular biology. Am J Clin Nutr63, 797S–811S.

    Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. 1951 Protein measurement with the Folin phenol reagent. J Biol Chem193, 265–275.

    Google Scholar 

  • Peña M, Lee J, Thiele D. 1999 A delicate balance: homeostatic control of copper uptake and distribution. J Nutr129, 1251–1260.

    Google Scholar 

  • Pinto M, Robine-Leon S, Appay M et al. 1983Enterocyte-like differentiation and polarization of the human colon carcinoma cell line Caco-2 in culture. Biol Cell47, 323–330.

    Google Scholar 

  • Ranaldi G, Islam K, Sambuy Y. 1994 D-cycloserine uses an active transport mechanism in the human intestinal cell line Caco-2. Antimicrob Agents Chemother38, 1239–1245.

    Google Scholar 

  • Reed D, Babson J, Beatty P, Brodie A, Ellis W, Potter D. 1980 High-performance liquid chromatography analysis of nanomole levels of glutathione, glutathione disulfide and related thiols and disulfides. Anal Biochem106, 55–62.

    Google Scholar 

  • Reeves P, Briske-Anderson M, Johnson L. 1998 Physiological concentrations of zinc affect the kinetics of copper uptake and transport in the human intestinal cell model, Caco-2. J Nutr128, 1794–1801.

    Google Scholar 

  • Reeves P, Briske-Anderson M, Newman S. 1996 High zinc concentrations in culture media affect copper uptake and transport in differentiated human colon adenocarcinoma cells. J Nutr126, 1701–1712.

    Google Scholar 

  • Rigo A, Rotilio G. 1977 Simultaneous determination of superoxide dismutase and catalase in biological materials by polarography. Anal Biochem81, 157–166.

    Google Scholar 

  • Rossi A, Poverini R, Di Lullo G, Modesti A, Modica A, Scarino ML. 1996 Heavy metal toxicity following apical and basolateral exposure in the human intestinal cell line Caco-2. Toxicol In Vitro10, 27–36.

    Google Scholar 

  • Schonewille J, Beynen A. 1995 High calcium intake does not impair apparent copper absorption in goats. J Anim Physiol Anim Nutr73, 251–257.

    Google Scholar 

  • Tandy S, Williams M, Legget A et al. 2000 Nramp2 expression is associated with pH-dependent iron uptake across the apical membrane of human intestinal Caco-2 cells. J Biol Chem275, 1023–1029.

    Google Scholar 

  • Tapia V, Arredondo M, Nuñez M. 1996 Regulation of Fe absorption by cultured intestinal epithelia (Caco-2) cell monolayers with varied Fe status. Am J Physiol271, G443–G447.

    Google Scholar 

  • van Campen D, Gross E. 1968 Influence of ascorbic acid on the absorption of copper by rats. J Nutr95, 617–622.

    Google Scholar 

  • van den Berg G, Yu S, Lemmens A, Beynen A. 1994 Dietary ascorbic acid lowers the concentration of soluble copper in the small intestineal lumen of rats. Br J Nutr71, 701–707.

    Google Scholar 

  • Wapnir R. 1998 Copper absorption and bioavailability. Am J Clin Nutr67 (suppl.), 1054S–1060S.

    Google Scholar 

  • Zhou B, Gitschier J. 1997 hCTR-1: a human gene for copper uptake identified by complementation in yeast. Proc Natl Acad Sci USA94, 7481–7486.

    Google Scholar 

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Ferruzza, S., Sambuy, Y., Ciriolo, M.R. et al. Copper uptake and intracellular distribution in the human intestinal Caco-2 cell line. Biometals 13, 179–185 (2000). https://doi.org/10.1023/A:1009271622356

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