Stereoselective binding of chiral ligands to single nucleotide polymorphisms of the human organic cation transporter-1 determined using cellular membrane affinity chromatography.
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.
Moaddel
R
R
Gerontology Research Center, National Institute on Aging, National Institutes of Health, Baltimore, MD 21224, USA. moaddelru@grc.nia.nih.gov
Bighi
F
F
Yamaguchi
R
R
Patel
S
S
Ravichandran
S
S
Wainer
I W
IW
D015655:1-Methyl-4-phenylpyridinium / Q000737:chemistry; D002460:Cell Line; D002462:Cell Membrane / Q000737:chemistry*; D002846:Chromatography, Affinity / Q000379:methods*; D005280:Fenoterol / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D007545:Isoproterenol / Q000737:chemistry / Q000378:metabolism; D008024:Ligands; D008958:Models, Molecular; D027702:Organic Cation Transporter 1 / Q000235:genetics*; D020641:Polymorphism, Single Nucleotide; D013237:Stereoisomerism; D014700:Verapamil / Q000737:chemistry / Q000378:metabolism
D008024:Ligands; D027702:Organic Cation Transporter 1; D005280:Fenoterol; D014700:Verapamil; D007545:Isoproterenol; D015655:1-Methyl-4-phenylpyridinium