Characterization of the MRP4- and MRP5-mediated transport of cyclic nucleotides from intact cells
| Autor: | Peter R. Wielinga, Piet Borst, Jan Wijnholds, Ingrid van der Heijden, Glen Reid, Jos H. Beijnen |
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| Rok vydání: | 2003 |
| Předmět: |
Nitroprusside
Ribosomal Proteins Time Factors Phosphodiesterase Inhibitors Phosphodiesterase 3 Blotting Western Nitric Oxide Biochemistry Cell Line chemistry.chemical_compound Cyclic nucleotide 1-Methyl-3-isobutylxanthine Cyclic AMP Humans Nitric Oxide Donors Cyclic nucleotide-gated ion channel Molecular Biology Cyclic GMP Forskolin Dose-Response Relationship Drug Biological Transport Cell Biology Glutathione Cyclic nucleotide transport chemistry CGMP transport Organic anion transport PDE10A Multidrug Resistance-Associated Proteins |
| Zdroj: | The Journal of biological chemistry. 278(20) |
| ISSN: | 0021-9258 |
| Popis: | Cyclic nucleotides are known to be effluxed from cultured cells or isolated tissues. Two recently described members of the multidrug resistance protein family, MRP4 and MRP5, might be involved in this process, because they transport the 3′,5′-cyclic nucleotides, cAMP and cGMP, into inside-out membrane vesicles. We have investigated cGMP and cAMP efflux from intact HEK293 cells overexpressing MRP4 or MRP5. The intracellular production of cGMP and cAMP was stimulated with the nitric oxide releasing compound sodium nitroprusside and the adenylate cyclase stimulator forskolin, respectively. MRP4- and MRP5-overexpressing cells effluxed more cGMP and cAMP than parental cells in an ATP-dependent manner. In contrast to a previous report we found no glutathione requirement for cyclic nucleotide transport. Transport increased proportionally with intracellular cyclic nucleotide concentrations over a calculated range of 20–600 μm, indicating low affinity transport. In addition to several classic inhibitors of organic anion transport, prostaglandins A1 and E1, the steroid progesterone and the anti-cancer drug estramustine all inhibited cyclic nucleotide efflux. The efflux mediated by MRP4 and MRP5 did not lead to a proportional decrease in the intracellular cGMP or cAMP levels but reduced cGMP by maximally 2-fold over the first hour. This was also the case when phosphodiesterase-mediated cyclic nucleotide hydrolysis was inhibited by 3-isobutyl-1-methylxanthine, conditions in which efflux was maximal. These data indicate that MRP4 and MRP5 are low affinity cyclic nucleotide transporters that may at best function as overflow pumps, decreasing steep increases in cGMP levels under conditions where cGMP synthesis is strongly induced and phosphodiesterase activity is limiting. |
| Databáze: | OpenAIRE |
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