Evers R, Kool M, van Deemter L, Janssen H, Calafat J, Oomen L C, Paulusma C C, Oude Elferink R P, Baas F, Schinkel A H, Borst P
Division of Molecular Biology, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands.
J Clin Invest. 1998 Apr 1;101(7):1310-9. doi: 10.1172/JCI119886.
The canalicular (apical) membrane of the hepatocyte contains an ATP-dependent transport system for organic anions, known as the multispecific organic anion transporter (cMOAT). The deduced amino acid sequence of cMOAT is 49% identical to that of the human multidrug resistance- associated protein (MRP) MRP1, and cMOAT and MRP1 are members of the same sub-family of adenine nucleotide binding cassette transporters. In contrast to MRP1, cMOAT was predominantly found intracellularly in nonpolarized cells, suggesting that cMOAT requires a polarized cell for plasma membrane routing. Therefore, we expressed cMOAT cDNA in polarized kidney epithelial MDCK cell lines. When these cells are grown in a monolayer, cMOAT localizes to the apical plasma membrane. We demonstrate that cMOAT causes transport of the organic anions S-(2,4-dinitrophenyl)-glutathione, the glutathione conjugate of ethacrynic acid, and S-(PGA1)-glutathione, a substrate not shown to be transported by organic anion transporters previously. Transport is inhibited only inefficiently by compounds known to block MRP1. We also show that cMOAT causes transport of the anticancer drug vinblastine to the apical side of a cell monolayer. We conclude that cMOAT is a 5'-adenosine triphosphate binding cassette transporter that potentially might be involved in drug resistance in mammalian cells.
肝细胞的胆小管(顶端)膜含有一种依赖ATP的有机阴离子转运系统,称为多特异性有机阴离子转运体(cMOAT)。cMOAT推导的氨基酸序列与人类多药耐药相关蛋白(MRP)MRP1的序列有49%的同源性,且cMOAT和MRP1是腺嘌呤核苷酸结合盒转运体同一家族的成员。与MRP1不同,cMOAT在非极化细胞中主要定位于细胞内,这表明cMOAT需要极化细胞来进行质膜转运。因此,我们在极化的肾上皮MDCK细胞系中表达了cMOAT cDNA。当这些细胞以单层生长时,cMOAT定位于顶端质膜。我们证明cMOAT能转运有机阴离子S-(2,4-二硝基苯基)-谷胱甘肽、依他尼酸的谷胱甘肽共轭物以及S-(PGA1)-谷胱甘肽,后者是一种以前未显示可被有机阴离子转运体转运的底物。已知能阻断MRP1的化合物对其转运的抑制作用较弱。我们还表明,cMOAT能将抗癌药物长春碱转运到细胞单层的顶端。我们得出结论,cMOAT是一种5'-三磷酸腺苷结合盒转运体,可能参与哺乳动物细胞的耐药性。