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Anal Biochem. 2010 Jun 1;401(1):148-53. doi: 10.1016/j.ab.2010.02.034. Epub 2010 Mar 3.

Stereoselective binding of chiral ligands to single nucleotide polymorphisms of the human organic cation transporter-1 determined using cellular membrane affinity chromatography.

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  • 1Gerontology Research Center, National Institute on Aging, National Institutes of Health, Baltimore, MD 21224, USA. moaddelru@grc.nia.nih.gov

Abstract

Membranes from stably transfected cell lines that express two point mutations of the human organic cation transporter-1 (hOCT1), R488M and G465R, have been immobilized on the immobilized artificial membrane (IAM) liquid chromatographic stationary phase to form two cellular membrane affinity chromatography (CMAC) columns, CMAC(hOCT1(G465R)) and CMAC(hOCT1(R488M)). Columns were created using both stationary phases, and frontal displacement chromatography experiments were conducted using [(3)H] MMP(+) (1-methyl-4-phenylpyridinium) as the marker ligand and various displacers, including the single enantiomers of verapamil, fenoterol, and isoproterenol. The chromatographic data obtained were used to refine a previously developed pharmacophore for hOCT1.

Published by Elsevier Inc.

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