tailieunhanh - Báo cáo khoa học: cGMP and glutathione-conjugate transport in human erythrocytes The roles of the multidrug resistance-associated proteins, MRP1, MRP4 and MRP5

The nature of cGMP transport in human erythrocytes, its relationship to glutathione conjugate transport, and pos-sible mediation by multidrug resistance-associated proteins (MRPs) have been investigated. MRP1, MRP4 and MRP5 are detected in immunoblotting studies with erythrocytes. MRP1and MRP5 are also detected in multidrug resistant COR-L23/R and MOR/R cells but at greatly reduced levels in the parent, drug sensitive COR-L23/P cells. MRP4 is detected in MOR/R but not COR-L23/R cells. | Eur. J. Biochem. 270 3696-3708 2003 FEBS 2003 doi cGMP and glutathione-conjugate transport in human erythrocytes The roles of the multidrug resistance-associated proteins MRP1 MRP4 and MRP5 Antonios Klokouzast Chung-Pu Wu Hendrik W. van Veen Margery A. Barrand and Stephen B. Hladky Department of Pharmacology University of Cambridge UK The nature of cGMP transport in human erythrocytes its relationship to glutathione conjugate transport and possible mediation by multidrug resistance-associated proteins MRPs have been investigated. MRP1 MRP4 and MRP5 are detected in immunoblotting studies with erythrocytes. MRP1and MRP5 are also detected in multidrug resistant COR-L23 R and MOR R cells but at greatly reduced levels in the parent drug sensitive COR-L23 P cells. MRP4 is detected in MOR R but not COR-L23 R cells. Uptake of cGMP into inside-out membrane vesicles prepared by a spontaneous one-step vesiculation process is shown to be by a low affinity system that accounts for more than 80 of the transport at all concentrations above 3 M. This transport is reduced by MRP inhibitors and substrates including MK-571 methotrexate estradiol 17-b-D-glucuronide and S 2 4-dinitrophenyl glutathione DNP-SG and also by glibenclamide and frusemide but not by the monoclonal Ig QCRL-3 that inhibits high-affinity transport of DNP-SG by MRP1. It is concluded that the cGMP exporter is distinct from MRP1and has properties similar to those reported for MRP4. Furthermore the evidence suggests that the protein responsible for cGMP transport is the same as that mediating low-affinity DNP-SG transport in human erythrocytes. Keywords multidrug resistance-associated protein guanosine cyclic mono-phosphate transport glutathione-conjugate transport human erythrocytes membrane vesicles. Active transport of the cyclic nucleotide cGMP across human erythrocyte membranes can be demonstrated using intact cells 1 or inside-out membrane vesicle preparations 2-4 . In the .

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