title: Mutation in an adaptor protein PDZK1 affects transport activity of organic cation transporter OCTNs and oligopeptide transporter PEPT2.
abstract: Genetic polymorphisms in xenobiotic transporters have recently been clarified to be associated with change in drug distribution and disposition. To expand on recent identification of direct interaction and functional regulation of several transporters by a PDZ (PSD95, Dlg and ZO1) domain containing protein PDZK1, the effect of mutation in PDZK1 on transport activity and subcellular localization of organic cation/carnitine transporters OCTN1 and OCTN2, and oligopeptide transporter PEPT2 was examined in the present study. HEK293 cells stably expressing a mutant transcript PDZK1-E195K (HEK293/PDZK1-E195K) were constructed, followed by transient transfection of cDNA for each transporter. Uptake of tetraethylammonium by OCTN1 was much higher in HEK293/PDZK1 cells, compared with that in the parent HEK293 cells, the uptake in HEK293/PDZK1-E195K cells showing middle range between the two values. Such difference in transport activity was accounted for the difference in transport capacity, with minimal change in affinity of OCTN1 to the substrate or other compounds. The similar difference among HEK293/PDZK1, HEK293/PDZK1-E195K and HEK293 cells was also observed in transport property of OCTN2 and PEPT2, whereas the difference was not so remarkable in each transporter with the last four amino acids deleted, that has much lower interaction potential with PDZK1. Immunohistochemical analysis indicated that OCTN1 was colocalized with PDZK1 on cell-surface, whereas colocalization with PDZK1-E195K was partially observed in cytoplasmic region. These results suggest a novel hypothesis that mutation in PDZK1 potentially changes transport property of various types of xenobiotic transporters by affecting their subcellular localization, possibly leading to change in disposition of various types of substrate drugs.
authors.affiliation: Division of Pharmaceutical Sciences, Graduate School of Natural Science and Technology, Kanazawa University, Kakuma-machi, Kanazawa, Japan.
authors.forename: Tomoko
authors.initials: T
authors.lastname: Sugiura
authors.forename: Yukio
authors.initials: Y
authors.lastname: Kato
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Kubo
authors.forename: Akira
authors.initials: A
authors.lastname: Tsuji
mesh_terms: D033982:Adaptor Protein Complex gamma Subunits / Q000378:metabolism; D001692:Biological Transport / Q000235:genetics / Q000502:physiology; D002250:Carbon Radioisotopes; D002331:Carnitine / Q000493:pharmacokinetics; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D002460:Cell Line; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D004151:Dipeptides / Q000493:pharmacokinetics; D006801:Humans; D008565:Membrane Proteins; D009154:Mutation / Q000235:genetics*; D027701:Organic Cation Transport Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D027702:Organic Cation Transporter 1 / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D011485:Protein Binding; D011738:Pyrilamine / Q000493:pharmacokinetics; D000074058:Solute Carrier Family 22 Member 5; D027981:Symporters / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D019789:Tetraethylammonium / Q000493:pharmacokinetics; D014162:Transfection; D014316:Tritium; D020798:Two-Hybrid System Techniques; D014700:Verapamil / Q000493:pharmacokinetics
chemical_list: D033982:Adaptor Protein Complex gamma Subunits; D002250:Carbon Radioisotopes; D002352:Carrier Proteins; D004151:Dipeptides; D008565:Membrane Proteins; D027701:Organic Cation Transport Proteins; D027702:Organic Cation Transporter 1; C484659:PDZK1 protein, human; C492020:SLC22A5 protein, human; D000074058:Solute Carrier Family 22 Member 5; D027981:Symporters; C093694:hydrogen-coupled oligopeptide transporter PepT2; D014316:Tritium; C004194:glycylsarcosine; D019789:Tetraethylammonium; D014700:Verapamil; D011738:Pyrilamine; D002331:Carnitine