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Vol. 53, Issue 6, 1068-1075, June 1998
Graduate School of Pharmaceutical Sciences (T.H., H.S., K.I., K.O.,
Y.S.) and
Graduate School of Medicine (K.K., T.S.) The University of
Tokyo, Hongo, Bunkyo-ku, Tokyo 113, Japan
The biliary excretion of several organic anions is mediated by the
canalicular multispecific organic anion transporter (cMOAT), which is
hereditarily defective in mutant rats such as Eisai hyperbilirubinemic rats (EHBR). In addition, using a kinetic study with isolated canalicular membrane vesicles, we recently suggested the presence of
ATP-dependent organic anion transporter(s) other than cMOAT in EHBR
[Pharm Res (NY) 12:1746-1755 (1995);
J Pharmacol Exp Ther 282:866-872 (1997)].
The aim of this study is to provide a molecular basis for the presence
of multiplicity in the biliary excretion of organic anions in rats.
Based on the homology with human multidrug resistance-associated
protein (hMRP), two cDNA fragments encoding the carboxyl-terminal
ATP-binding cassette region were amplified by reverse
transcription-polymerase chain reaction from EHBR liver. These
fragments exhibited approximately 70% amino acid identity with hMRP
and rat cMOAT;, therefore, they were designated MRP-like proteins
(MLP-1 and MLP-2). The cloned full length cDNA of MLP-1 and -2 from the
Sprague-Dawley (SD) rat liver and colon cDNA library was composed of
1502 and 1523 amino acids, respectively, had the characteristics of
ATP-binding cassette transporters, and exhibited homology with hMRP and
rat cMOAT. Northern blot analysis indicated that MLP-1 is expressed predominantly in the liver in both SD rats and EHBR, whereas hepatic expression of MLP-2 was observed only in EHBR. In addition, MLP-2 was
markedly induced by ligation of the bile duct in SD rat liver. In both
SD rats and EHBR, MLP-2 was expressed predominantly in the duodenum,
jejunum, and colon. These findings suggest that MLP-1 and MLP-2 might
be novel members of the MRP family responsible for the excretion of
organic anions from these epithelial cells, and that MLP-2 is an
inducible one.
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