title: Primary structure and functional expression of the apical organic cation transporter from kidney epithelial LLC-PK1 cells.
abstract: Renal secretion of organic cations involves at least two distinct transporters, located in the basolateral and apical membranes of proximal tubule cells. Whereas the basolateral transporter has recently been cloned, sequence information about the apical type was not yet available. An organic cation transporter, OCT2p, was cloned from LLC-PK1 cells, a porcine cell line with properties of proximal tubular epithelial cells. OCT2p was heterologously expressed and characterized in human embryonic kidney 293 cells. OCT2p-mediated uptake of the prototypical organic cation [14C]tetraethylammonium ([14C]TEA) into 293 cells was saturable. There was a highly significant correlation between the Ki values for the inhibition of apical [14C]TEA uptake into LLC-PK1 cells and 293 cells transfected with OCT2p (r = 0.995; p < 0.001; n = 6). Although OCT2p is structurally related to OCT1r, the basolateral organic cation transporter from rat kidney, the transporters could be clearly discriminated pharmacologically with corticosterone, decynium22, and O-methylisoprenaline. The findings at hand suggest that OCT2 corresponds to the apical type of organic cation transporter. Reverse transcriptase-polymerase chain reaction indicates that mRNA of OCT1r is limited to non-neuronal tissue, whereas OCT2r, the OCT2p homologue from rat, was found in both the kidney and central nervous regions known to be rich in the monoamine transmitter dopamine.
authors.affiliation: Department of Pharmacology, University of Heidelberg, Im Neuenheimer Feld 366, 69120 Heidelberg, Germany. dirk.gruendemann@urz.uni-heidelberg.de
authors.forename: D
authors.initials: D
authors.lastname: Gründemann
authors.forename: J
authors.initials: J
authors.lastname: Babin-Ebell
authors.forename: F
authors.initials: F
authors.lastname: Martel
authors.forename: N
authors.initials: N
authors.lastname: Ording
authors.forename: A
authors.initials: A
authors.lastname: Schmidt
authors.forename: E
authors.initials: E
authors.lastname: Schömig
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001692:Biological Transport; D002352:Carrier Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000502:physiology*; D002490:Central Nervous System / Q000502:physiology; D016384:Consensus Sequence; D017931:DNA Primers; D006801:Humans; D007668:Kidney / Q000378:metabolism*; D018374:LLC-PK1 Cells; D008961:Models, Structural; D008969:Molecular Sequence Data; D009928:Organ Specificity; D027701:Organic Cation Transport Proteins; D000075102:Organic Cation Transporter 2; D016133:Polymerase Chain Reaction; D011487:Protein Conformation; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D013552:Swine; D019789:Tetraethylammonium; D013757:Tetraethylammonium Compounds / Q000493:pharmacokinetics; D014162:Transfection
chemical_list: D002352:Carrier Proteins; D017931:DNA Primers; D027701:Organic Cation Transport Proteins; D000075102:Organic Cation Transporter 2; D011994:Recombinant Proteins; C491013:SLC22A2 protein, human; C100590:Slc22a2 protein, rat; D013757:Tetraethylammonium Compounds; D019789:Tetraethylammonium