Interactions of mefloquine with ABC proteins, MRP1 (ABCC1) and MRP4 (ABCC4) that are present in human red cell membranes
Introduction
Multidrug resistant-associated proteins (MRPs) mediate ATP-dependent transport of a wide range of compounds including glutathione conjugates and cyclic nucleotides. MRP1 (ABCC1) is a primary active transporter of organic anions including the oxidised form of glutathione, GSSG [1]. It is expressed in human erythrocytes where it accounts for the high affinity component of the transport of GSSG and dinitrophenyl-S-glutathione, DNP-SG [2], [3], [4]. In addition to MRP1 there are in red blood cells other efflux transporters, one or more of which account for the low affinity efflux of GSSG, DNP-SG and cGMP. The identities of these transporters have not been conclusively identified though MRP4 is thought to play a major role [4]. One of the most potent inhibitors of MRP-mediated transport is the LTD4 receptor antagonist MK-571, a quinoline derivative. Another quinoline-based compound, IAAQ, photolabels MRP1 and both chloroquine and MK-571 inhibit this labelling [5]. Thus it is probable that at least some quinolines in clinical use interact with MRPs. Quinoline-based antimalarials, especially chloroquine, have been used for decades to kill malaria parasites in the intraerythrocytic stage of their life cycle, If indeed these quinolines do interact with active transporters on erythrocytes, the possibility exists that they could affect access of drug to the parasite inside the cell. Interaction of mefloquine with P-glycoprotein, a mammalian ATP-binding cassette transporter has been documented [6], [7] but, while it has been sought, P-glycoprotein has not been detected in human erythrocyte membranes by immunoblotting [8] (Wu and Woodcock personal communication). Little or no information is available regarding the interaction of mefloquine with other ABC transporters such as the MRPs that are present in red blood cells.
The aim of the present study was to explore how some widely used quinoline-based antimalarial drugs affect activities of some MRPs known to be present in human erythrocytes. Using inside-out vesicles prepared from human red blood cells, we show that MK-571 and mefloquine directly inhibit MRP-mediated uptake of DNP-SG or cGMP. Confirmation of these interactions was obtained using inside-out vesicles prepared from MRP1-overexpressing tumour cells or transfected HEK 293 cells overexpressing MRP4. Further evidence of their interactions was obtained by observing their effects on accumulation and efflux of MRP substrates at the whole cell level. The effects of quinolines on ATPase activities and ATP binding to MRP1 and MRP4 protein provide further insight into the nature of the quinoline-MRP interactions and suggest that MK-571 and mefloquine appear to be substrates for transport.
Section snippets
Cell culture
Cells of the human large-cell lung tumour line L23/P and of its doxorubicin-selected MRP1-overexpressing multidrug-resistant variant L23/R [9], [10] were cultured as previously described [4]. Parental 293 human embryonic kidney cells (HEK 293 cells) and the MRP4-overexpressing HEK 293/4.63 cells [11] were generous gifts of Prof P. Borst (Division of Molecular Biology and Center for Biomedical Genetics, The Netherlands Cancer Institute, Amsterdam, The Netherlands). HEK 293/4.63 cells were
Effects of quinoline derivatives on high affinity DNP-SG transport in human erythrocyte vesicles
ATP-dependent uptake of DNP-SG was measured over the course of 30 min in the absence or presence of quinolines. It has previously been shown [31] that DNP-SG uptake into human erythrocyte vesicles is linear for at least 60 min. The rate of uptake was significantly inhibited by MK-571 and by mefloquine with IC50 values of 1.1 ± 0.2 μM (n = 7, maximum percentage uptake 94 ± 4%) and 127 ± 17 μM (n = 4, maximum percentage uptake 98 ± 3%), respectively (see Fig. 1A). Other quinoline antimalarial drugs (amodiaquine,
Discussion
The purpose of this study was to investigate the interactions of mefloquine and other quinolines used in the treatment of malaria with MRPs present in red blood cell membranes. Of these, mefloquine was the most effective. It blocked the higher affinity DNP-SG transport observed in red blood cells and the MRP1-mediated DNP-SG transport in lung tumour cells as well as the principal cGMP transport in red blood cells and the cGMP transport measured in MRP4-transfected HEK cells with IC50 values of
Acknowledgements
We thank Dr. Gary Kruh (Fox chase Cancer Centre) for the gifts of plasmid PVL1393-MRP4 and anti-MRP4 and Dr. Piet Borst (The Netherlands Cancer Institute) for HEK 294/4.63 cells. CPW is supported by the Cambridge Commonwealth Trust Fund and by a visiting preCRTA award from the NCI, NIH.
References (47)
- et al.
Direct binding of chloroquine to the multidrug resistance protein (MRP). Possible role for MRP in chloroquine drug transport and resistance in tumor cells
Biochem Pharmacol
(1998) - et al.
Modulation of the function of human MDR1 P-glycoprotein by the antimalarial drug mefloquine
Biochem Pharmacol
(1996) - et al.
Mutually co-operative interactions between modulators of p-glycoprotein
Biochim Biophys Acta: Mol Basis Dis
(1997) - et al.
Cellular and biophysical evidence for interactions between adenosine triphosphate and P-glycoprotein substrates: functional implications for adenosine triphosphate/drug cotransport in P-glycoprotein overexpressing tumor cells and in P-glycoprotein low-level expressing erythrocytes
Blood Cells Mol Dis
(2001) - et al.
GS-X pump is functionally overexpressed in cis-diamminedichloroplatinum(ii)-resistant human leukemia HL-60 cells and down-regulated by cell-differentiation
J Biol Chem
(1994) - et al.
Influences of glutathione on anionic substrate efflux in tumour cells expressing the multidrug resistance-associated protein, MRP1
Biochem Pharmacol
(2001) - et al.
Correlation between steady-state ATP hydrolysis and vanadate-induced ADP trapping in human P-glycoprotein—evidence for ADP release as the rate-limiting step in the catalytic cycle and its modulation by substrates
J Biol Chem
(2001) - et al.
A rapid, sensitive, and specific method for the determination of protein in dilute solution
Anal Biochem
(1973) - et al.
Functionally similar vanadate-induced 8-azidoadenosine 5′-alpha-P-32 diphosphate-trapped transition state intermediates of human P-glycoprotein are generated in the absence and presence of ATP hydrolysis
J Biol Chem
(2001) Drug-stimulatable ATPase activity in crude membranes of human MDR1-transfected mammalian cells
Meth Enzymol
(1998)
Characterization of the catalytic cycle of ATP hydrolysis by human P-glycoprotein—the two ATP hydrolysis events in a single catalytic cycle are kinetically similar but affect different functional outcomes
J Biol Chem
ATP/Mg2+-dependent cardiac transport system for glutathione S-conjugates. A study using rat heart sarcolemma vesicles
J Biol Chem
Enzymatic conjugation of erythrocyte glutathione with 1-chloro-2,4-dintrobenzene: the fate of glutathione conjugate in ertythrocytes and the effect of glutathione depletion on hemoglobin
Blood
Characterization of the MRP4- and MRP5-mediated transport of cyclic nucleotides from intact cells
J Biol Chem
Multidrug resistance protein 4 (MRP4/ABCC4)-mediated ATP hydrolysis: Effect of transporty substrates and characterization of the post-hydrolysis transition state
J Biol Chem
Relation between the turnover number for vinblastine transport and for vinblastine-stimulated ATP hydrolysis by human P-glycoprotein
J Biol Chem
ATPase activity of purified multidrug resistance-associated protein
J Biol Chem
ATPase activity of purified and reconstituted multidrug resistance protein MRP1 from drug-selected H69AR cells
Biochim Biophys Acta: Biomembr
Plasmodium falciparum expresses a multidrug resistance associated protein
Biochem Biophys Res Commun
The multidrug resistance protein 5 functions as an ATP-dependent export pump for cyclic nucleotides
J Biol Chem
ATP-dependent glutathione disulphide transport mediated by the MRP gene-encoded conjugate export pump
Biochem J
Identification of the multidrug-resistance protein (MRP) as the glutathione-S-conjugate export pump of erythrocytes
Eur J Biochem
Increased sensitivity to anticancer drugs and decreased inflammatory response in mice lacking the multidrug resistance-associated protein
Nat Med
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