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Originally published In Press as doi:10.1074/jbc.M104833200 on July 10, 2001

J. Biol. Chem., Vol. 276, Issue 36, 33747-33754, September 7, 2001
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Transport of Cyclic Nucleotides and Estradiol 17-beta -D-Glucuronide by Multidrug Resistance Protein 4
RESISTANCE TO 6-MERCAPTOPURINE AND 6-THIOGUANINE*

Zhe-Sheng ChenDagger §, Kun LeeDagger , and Gary D. Kruh||

From the Medical Science Division, Fox Chase Cancer Center, Philadelphia, Pennsylvania 19111

Human multidrug resistance protein 4 (MRP4) has recently been determined to confer resistance to the antiviral purine analog 9-(2-phosphonylmethoxyethyl)adenine and methotrexate. However, neither its substrate selectivity nor physiological functions have been determined. Here we report the results of investigations of the in vitro transport properties of MRP4 using membrane vesicles prepared from insect cells infected with MRP4 baculovirus. It is shown that expression of MRP4 is specifically associated with the MgATP-dependent transport of cGMP, cAMP, and estradiol 17-beta -D-glucuronide (E217beta G). cGMP, cAMP, and E217beta G are transported with Km and Vmax values of 9.7 ± 2.3 µM and 2.0 ± 0.3 pmol/mg/min, 44.5 ± 5.8 µM and 4.1 ± 0.4 pmol/mg/min, and 30.3 ± 6.2 µM and 102 ± 16 pmol/mg/min, respectively. Consistent with its ability to transport cyclic nucleotides, it is demonstrated that the MRP4 drug resistance profile extends to 6-mercaptopurine and 6-thioguanine, two anticancer purine analogs that are converted in the cell to nucleotide analogs. On the basis of its capacity to transport cyclic nucleotides and E217beta G, it is concluded that MRP4 may influence diverse cellular processes regulated by cAMP and cGMP and that its substrate range is distinct from that of any other characterized MRP family member.

* This work was supported in part by National Institutes of Health Grant CA73728 (to G. D. K.) and by an appropriation from the Commonwealth of Pennsylvania.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger Contributed equally to this work.

§ Recipient of a Japan Research Foundation for Clinical Pharmacology award.

Recipient of National Institutes of Health Fellowship CA74518.

|| To whom correspondence should be addressed: Fox Chase Cancer Center, 7701 Burholme Ave., Philadelphia, PA 19111. Tel.: 215-728-5317; Fax: 215-728-3603; E-mail: GD_Kruh@fccc.edu.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.


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