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Cloning and heterologous expression of the ovine (Ovis aries) P-glycoprotein (Mdr1) in Madin-Darby canine kidney (MDCK) cells

2010; Wiley; Volume: 33; Issue: 3 Linguagem: Inglês

10.1111/j.1365-2885.2009.01141.x

1365-2885

David Zahner, Julia M. Alber, Ernst Petzinger,

Drug-Induced Hepatotoxicity and Protection

Zahner, D., Alber, J., Petzinger, E. Cloning and heterologous expression of the ovine (Ovis aries) P-glycoprotein (Mdr1) in Madin–Darby canine kidney (MDCK) cells. J. vet. Pharmacol. Therap.33, 304–311. P-glycoprotein (P-gp) plays a crucial role in the multidrug resistance of pathogenic helminths in sheep (Ovis aries) as well as in antiparasitic drug pharmacokinetics in the host. We cloned sheep P-gp cDNA and expressed it stably in Madin–Darby canine kidney (MDCK) cells. The open reading frame consists of 3858 nucleotides coding for a 1285 amino acids containing protein. The sequence shows high homology to the orthologs of other mammalian species, especially cattle. Both ruminant DNA sequences show a 9 bp insertion that is lacking in all other investigated sequences. Expressed in MDCK cells, the protein displays a size of 170 kDa on Western analysis. Transfection of MDCK cells with sheep P-gp resulted in 10- to 50-fold resistance to the cytotoxic P-gp substrates colchicin and daunorubicin, and in reduced digoxin accumulation.