title: Steroid and bile acid conjugates are substrates of human multidrug-resistance protein (MRP) 4 (ATP-binding cassette C4).
abstract: Human multidrug-resistance protein (MRP) 4 transports cyclic nucleotides and when overproduced in mammalian cells mediates resistance to some nucleoside analogues. Recently, it has been shown that Mrp4 is induced in the livers of Fxr ((-/-)) mice, which have increased levels of serum bile acids. Since MRP4, like MRP1-3, also mediates transport of a model steroid conjugate substrate, oestradiol 17-beta-D-glucuronide (E(2)17betaG), we tested whether MRP4 may be involved in the transport of steroid and bile acid conjugates. Bile salts, especially sulphated derivatives, and cholestatic oestrogens inhibited the MRP4-mediated transport of E(2)17betaG. Inhibition by oestradiol 3,17-disulphate and taurolithocholate 3-sulphate was competitive, suggesting that these compounds are MRP4 substrates. Furthermore, we found that MRP4 transports dehydroepiandrosterone 3-sulphate (DHEAS), the most abundant circulating steroid in humans, which is made in the adrenal gland. The ATP-dependent transport of DHEAS by MRP4 showed saturable kinetics with K (m) and V (max) values of 2 microM and 45 pmol/mg per min, respectively (at 27 degrees C). We further studied the possible involvement of other members of the MRP family of transporters in the transport of DHEAS. We found that MRP1 transports DHEAS in a glutathione-dependent manner and exhibits K (m) and V (max) values of 5 microM and 73 pmol/mg per min, respectively (at 27 degrees C). No transport of DHEAS was observed in membrane vesicles containing MRP2 or MRP3. Our findings suggest a physiological role for MRP1 and MRP4 in DHEAS transport and an involvement of MRP4 in transport of conjugated steroids and bile acids.
authors.affiliation: Division of Molecular Biology and Center for Biomedical Genetics, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands.
authors.forename: Noam
authors.initials: N
authors.lastname: Zelcer
authors.forename: Glen
authors.initials: G
authors.lastname: Reid
authors.forename: Peter
authors.initials: P
authors.lastname: Wielinga
authors.forename: Annemieke
authors.initials: A
authors.lastname: Kuil
authors.forename: Ingrid
authors.initials: I
authors.lastname: van der Heijden
authors.forename: John D
authors.initials: JD
authors.lastname: Schuetz
authors.forename: Piet
authors.initials: P
authors.lastname: Borst
mesh_terms: D001647:Bile Acids and Salts / Q000378:metabolism*; D019314:Dehydroepiandrosterone Sulfate / Q000378:metabolism; D018432:Drug Resistance, Multiple; D004958:Estradiol / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D013256:Steroids / Q000378:metabolism*; D013379:Substrate Specificity; D014162:Transfection
chemical_list: D001647:Bile Acids and Salts; D019314:Dehydroepiandrosterone Sulfate; D004958:Estradiol; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D011994:Recombinant Proteins; D013256:Steroids; C073492:ABCC4 protein, human