Molecular cloning and pharmacological characterization of rat multidrug resistance protein 1 (mrp1).
Multidrug resistance protein 1 (MRP1) transports a wide range of structurally diverse conjugated and nonconjugated organic anions and some peptides, including oxidized and reduced glutathione (GSH). The protein confers resistance to certain heavy metal oxyanions and a variety of natural product-type chemotherapeutic agents. Elevated levels of MRP1 have been detected in many human tumors, and the protein is a candidate therapeutic target for drug resistance reversing agents. Previously, we have shown that human MRP1 (hMRP1) and murine MRP1 (mMRP1) differ in their substrate specificity despite a high degree of structural conservation. Since rat models are widely used in the drug discovery and development stage, we have cloned and functionally characterized rat MRP1 (rMRP1). Like mMRP1 and in contrast to hMRP1, rMRP1 confers no, or very low, resistance to anthracyclines and transports the two estrogen conjugates, 17beta-estradiol-17-(beta-d-glucuronide) (E217betaG) and estrone 3-sulfate, relatively poorly. Mutational studies combined with vesicle transport assays identified several amino acids conserved between rat and mouse, but not hMRP1, that make major contributions to these differences in substrate specificity. Despite the fact that the rodent proteins transport E217betaG poorly and the GSH-stimulated transport of estrone 3-sulfate is low compared with hMRP1, site-directed mutagenesis studies indicate that different nonconserved amino acids are involved in the low efficiency with which each of the two estrogen conjugates is transported. Our studies also suggest that although rMRP1 and mMRP1 are 95% identical in primary structure, their substrate specificities may be influenced by amino acids that are not conserved between the two rodent proteins.
Nunoya
Kenichi
K
Department of Xenobiotic and Disposition, Minase Research Institute, Ono Pharmaceutical Co, Ltd, OSaka, Japan.
Grant
Caroline E
CE
Zhang
Dawei
D
Cole
Susan P C
SP
Deeley
Roger G
RG
D000595:Amino Acid Sequence / Q000235:genetics; D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism; D002460:Cell Line; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004317:Doxorubicin / Q000378:metabolism / Q000493:pharmacokinetics; D004622:Embryo, Mammalian; D004958:Estradiol / Q000031:analogs & derivatives / Q000378:metabolism; D045166:Estradiol Congeners / Q000378:metabolism; D004970:Estrone / Q000031:analogs & derivatives* / Q000037:antagonists & inhibitors / Q000378:metabolism; D005786:Gene Expression Regulation / Q000235:genetics; D006801:Humans; D007668:Kidney / Q000166:cytology; D017997:Leukotriene C4 / Q000378:metabolism; D051379:Mice / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000493:pharmacokinetics*; D016297:Mutagenesis, Site-Directed; D012333:RNA, Messenger; D051381:Rats / Q000235:genetics / Q000378:metabolism*; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D013379:Substrate Specificity / Q000502:physiology; D014162:Transfection; D014316:Tritium
D002352:Carrier Proteins; D018076:DNA, Complementary; D004317:Doxorubicin; D004958:Estradiol; D045166:Estradiol Congeners; D004970:Estrone; D017997:Leukotriene C4; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D012333:RNA, Messenger; D014316:Tritium; C025483:estradiol-17 beta-glucuronide; C017296:estrone sulfate