Summary
The characteristics of uridine transport were studied in basolateral plasma membrane vesicles isolated from rat liver. Uridine was not metabolized under transport measurement conditions and was taken up into an osmotically active space with no significant binding of uridine to the membrane vesicles. Uridine uptake was sodium dependent, showing no significant stimulation by other monovalent cations. Kinetic analysis of the sodium-dependent component showed a single system with Michaelis-Menten kinetics. Parameter values were K M 8.9 μ m and V max 0.57 pmol/mg prot/sec. Uridine transport proved to be electrogenic, since, firstly, the Hill plot of the kinetic data suggested a 1 uridine: 1 Na+ stoichiometry, secondly, valinomycin enhanced basal uridine uptake rates and, thirdly, the permeant nature of the Na+ counterions determined uridine transport rates (SCN− > NO −3 > Cl− > SO 2−4 ). Other purines and pyrimidines cis-inhibited and trans-stimulated uridine uptake.
Similar content being viewed by others
References
Barros, L.F., Bustamante, J.C., Yudilevich, D.L., Jarvis, S.M. 1991. Adenosine transport and nitrobenzylthioinosine binding in human placental membrane vesicles from brush-border and basal sides of the trophoblast. J. Membrane Biol. 119:151–161
Bartrons, R., Van Schaftingen, E., Hers, H.-G. 1984. The ability of adenosine to decrease the concentration of fructose-2,6-bisphosphate in isolated hepatocytes. Biochem. J. 218:157–163
Belt, J.A., Noel, L.D. 1985. Nucleoside transport in Walker 256 rat carcino sarcoma and S49 mouse lymphoma cells. Biochem. J. 232:681–688
Buxton, D.B., Fisher, R.A., Robertson, S.M., Olson, M.S. 1987. Stimulation of glycogenolysis and vasoconstriction by adenosine and adenosine analogues in the perfused rat liver. Biochem. J. 248:35–41
Casado, J., Felipe, A., Pastor-Anglada, M., Remesar, X. 1988. Glutamine as a major nitrogen carrier to the liver in suckling rat pups. Biochem. J. 256:377–381
Dagnino L., Bennet, L.L., Paterson, A.R.P. 1991. Substrate specificity, kinetics, and stoichiometry of sodium-dependent adenosine transport in L1210/AM mouse leukemia cells. J. Biol. Chem. 266:6312–6317
Dagnino, L., Paterson, A.R.P. 1990. Sodium-dependent and equilibrative nucleoside transport systems in L1210 mouse leukemia cells: Effect of inhibitors of equilibrative systems on the content and retention of nucleosides. Cancer Res. 50:6549–6553
Felipe, A., Remesar, X., Pastor-Anglada, M. 1989. Na+-dependent alanine transport in plasma membrane vesicles from late-pregnant rat livers. Pediatr. Res. 26:448–451
Gasser, T., Moyer, J.D., Handschumacher, R.E. 1981. Novel single-pass exchange of circulating uridine in rat liver. Science 213:777–778
Guinzberg, P., Laguna, I., Zentella, A., Guzman, R., Piña, E. 1987. Effect of adenosine and inosine on ureagenesis in hepatocytes. Biochem. J. 245:371–374
Holstege, A., Gengenbacher, H.-M., Jehle, L., Hoppmann, J. 1991. Facilitated diffusion and sodium-dependent transport of purine and pyrimidine nucleosides in rat liver. Hepatology 14:373–380
Holstege, A., Manglitz, D., Gerok, W. 1984. Depletion of blood plasma cytidine due to increase hepatocellular salvage in d-galactosamine-treated rats. Eur. J. Biochem. 141:339–344
Jakobs, E.S., Van Os-Corby, D.J., Paterson, A.R.P. 1990. Expression of sodium-linked nucleoside transport activity in monolayer cultures of IEC-6 intestinal epithelial cells. J. Biol. Chem. 265:22210–22216
Jarvis, S.M. 1988. Inhibition by nucleosides of glucose-transport activity in human erythrocytes. Biochem. J. 249:383–389
Jarvis, S.M. 1989. Characterization of sodium-dependent nu-cleoside transport in rabbit intestinal brush-border membrane vesicles. Biochim. Biophys. Acta 979:132–138
Jarvis, S.M., Griffith, D.A. 1991. Expression of the rabbit intestinal N2 Na+/nucleoside transporter in Xenopus laevis oocytes. Biochem. J. 278:605–607
Lavoinne, A., Claeyssens, S., Chedeville, A. 1990. Metabolism of adenosine through adenosine kinase inhibits gluconeogenesis in isolated rat hepatocytes. Eur. J. Biochem. 187:403–407
Lee, C.W., Cheeseman, C.I., Jarvis, S.M. 1988. Na+-and K+-dependent uridine transport in rat renal brush-border membrane vesicles. Biochim. Biophys. Acta 942:139–149
May, J.M. 1988. Inhibition of hexose transport by adenosine derivatives in human erythrocytes. J. Cell. Physiol. 135:332–338
Möser, G.H., Schrader, J., Deussen, A. 1989. Turnover of adenosine in plasma of human and dog blood. Am. J. Physiol. 256:C799-C806
Oetjen, E., Schweickhardt, C., Unthan-Fechner, K., Probst, I. 1990. Stimulation of glucose production from glycogen by glucagon, noradrenaline and non-degradable adenosine analogues is counteracted by adenosine and ATP in cultured rat hepatocytes. Biochem. J. 271:337–344
Ontyd, J., Schrader, J. 1984. Measurement of adenosine, inosine, hypoxanthine in human plasma. J. Chromatogr. 307:404–409
Pastor-Anglada, M., Remesar, X., Bourdel, G. 1987. Alanine uptake by liver at midpregnancy in rat. Am. J. Physiol. 252:E408-E413
Peterson, G.L. 1979. Review of the Folin phenol protein quantitation method of Lowry, Farr and Randall. Anal. Biochem. 100:201–220
Plagemann, P.G.W. 1991. Na+-dependent, concentrative nucleoside transport in rat macrophages. Specificity for natural nucleosides and nucleoside analogs, including dideoxy-nucleosides, and comparison of nucleoside transport in rat, mouse and human macrophages. Biochem. Pharmacol. 42:247–252
Plagemann, P.G.W., Aran, J.M. 1990. Na+-dependent, active nucleoside transport in mouse spleen lymphocytes, leukemia cells, fibroblasts and macrophages, but not in equivalent human or pig cells; dipyridamole enhances nucleoside salvage by cells with both active and facilitated transport. Biochim. Biophys. Acta 1025:32–42
Plagemann, P.G.W., Aran, J.M. 1990. Characterization of Na+-dependent, active nucleoside transport in rat and mouse peritoneal macrophages, a mouse macrophage cell line and normal rat kidney cells. Biochim. Biophys. Acta 1028:289–298
Plagemann, P.G.W., Woffendin, C. 1989. Na+-dependent and -independent transport of uridine and its phosphorylation in mouse spleen cells. Biochim. Biophys. Acta 981:315–325
Plagemann, P.G.W., Wohlhueter, R.M. 1984. Nucleoside transport in cultured mammalian cells. Multiple forms with different sensitivity to inhibition by nitrobenzylthionosine or hypoxanthine. Biochim. Biophys. Acta 773:39–52
Plagemann, P.G.W., Wohlhueter, R.M. 1985. Nitrobenzyl-thioinosine-sensitive and -resistant nucleoside transport in normal and transformed rat cells. Biochim. Biophys. Acta 816:387–395
Plagemann, P.G.W., Wohlhueter, R.M., Woffendin, C. 1988. Nucleoside and nucleobase transport in animal cells. Biochim. Biophys. Acta 947:405–443
Quintana, I., Felipe, A., Remesar, X., Pastor-Anglada, M. 1988. Carrier-mediated uptake of l-(+)-lactate in plasma membrane vesicles from rat liver. FEBS Lett. 235:224–228
Ruiz, B., Felipe, A., Casado, J., Pastor-Anglada, M. 1991. Amino acid uptake by livers of genetically obese Zucker rats. Biochem. J. 280:367–372
Schwenk, M., Hegazy, E., Lopez del Pino V. 1984. Uridine uptake by isolated intestinal epithelial cells of guinea pig. Biochim. Biophys. Acta 805:370–374
Van Amelsvoort, J.M.M., Sips, H.J., Van Dam, K. 1978. Sodium-dependent alanine transport in plasma-membrane vesicles from rat liver. Biochem. J. 174:1083–1086
Williams, T.C., Doherty, A., Griffith, D.A., Jarvis, S.M. 1989. Characterization of sodium-dependent and sodium-independent nucleoside transport systems in rabbit brush-border and basolateral plasma-membrane vesicles from the renal outer cortex. Biochem. J. 264:223–231
Williams, T.C., Jarvis, S.M. 1991. Multiple sodium-dependent nucleoside transport systems in bovine renal brush-border membrane vesicles. Biochem. J. 274:27–33
Wu, J.-S.R., Kwong, F.Y.P., Jarvis, S.M., Young, J.D. 1983. Identification of the erythrocyte nucleoside transporter as a band 4.5 polypeptide. Photoaffinity labelling studies using nitrobenzylthioinosine. J. Biol. Chem. 258:13745–13751
Wu, J.-S.R., Young, J.D. 1984. Photoaffinity labeling of nucleoside-transport proteins in plasma membranes isolated from rat and guinea-pig liver. Biochem. J. 220:499–506
Author information
Authors and Affiliations
Additional information
This work has been partially supported by grant PM90-0162 from D.G.I.C.Y.T. (Ministerio de Educación y Ciencia, Spain). B.R.-M. is a research fellow supported by the Nestlé Nutrition Research Grant Programme.
Rights and permissions
About this article
Cite this article
Ruiz-Montasell, B., Casado, F.J., Felipe, A. et al. Uridine transport in basolateral plasma membrane vesicles from rat liver. J. Membarin Biol. 128, 227–233 (1992). https://doi.org/10.1007/BF00231815
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00231815