Role of rat organic anion transporter 3 (Oat3) in the renal basolateral transport of glutathione.
The tripeptide GSH is important in maintenance of renal redox status and defense against reactive electrophiles and oxidants. Previous studies showed that GSH is transported across the basolateral plasma membrane (BLM) into the renal proximal tubule by both sodium-coupled and sodium-independent pathways. Substrate specificity and inhibitor studies suggested the function of several carriers, including organic anion transporter 3 (Oat3). To test the hypothesis that rat Oat3 can function in renal GSH transport, the cDNA for rat Oat3 was expressed as a His6-tagged protein in E. coli, purified from inclusion bodies and by Ni2+-affinity chromatography, and reconstituted into proteoliposomes. cDNA-expressed and reconstituted Oat3 transported both GSH and p-aminohippurate (PAH) in exchange for 2-oxoglutarate (2-OG) and 2-OG and PAH in exchange for GSH, and PAH uptake was inhibited by both probenecid and furosemide, consistent with function of Oat3. mRNA expression of Oat3 and several other potential carriers was detected by RT-PCR in rat kidney cortex but was absent from NRK-52E cells, a rat proximal tubular cell line. Basolateral uptake of GSH in NRK-52E cells showed little PAH- or 2-OG-stimulated uptake. We conclude that Oat3 can function in GSH uptake and that NRK-52E cells possess a low background rate of GSH uptake, making these cells a good model for overexpression of specific, putative GSH carriers.
Lash
Lawrence H
LH
Department of Pharmacology, Wayne State University School of Medicine, 540 East Canfield Avenue, Detroit, MI 48201, United States. l.h.lash@wayne.edu <l.h.lash@wayne.edu>
Putt
David A
DA
Xu
Feng
F
Matherly
Larry H
LH
D000818:Animals; D001483:Base Sequence; D001485:Basement Membrane / Q000378:metabolism; D001692:Biological Transport; D002460:Cell Line; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D005978:Glutathione / Q000378:metabolism*; D007672:Kidney Cortex / Q000166:cytology / Q000378:metabolism*; D029362:Organic Anion Transporters, Sodium-Independent / Q000502:physiology*; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction
D017931:DNA Primers; D029362:Organic Anion Transporters, Sodium-Independent; D012333:RNA, Messenger; C114338:organic anion transport protein 3; D005978:Glutathione
Annu Rev Physiol. 1999;61:663-82
10099705
J Pharmacol Exp Ther. 1998 May;285(2):608-18
9580605
Toxicol Appl Pharmacol. 2005 May 1;204(3):329-42
15845422
J Pharmacol Exp Ther. 2006 Mar;316(3):1175-86
16291728
Arch Biochem Biophys. 2000 Jan 1;373(1):193-202
10620338
Am J Physiol Renal Physiol. 2000 Aug;279(2):F216-32
10919840
Pflugers Arch. 2000 Jul;440(3):337-50
10954321
Kidney Int. 2001 Jan;59(1):17-30
11135053
News Physiol Sci. 2001 Jun;16:114-8
11443229
Mol Pharmacol. 2001 Nov;60(5):1091-9
11641438
Annu Rev Physiol. 2002;64:563-94
11826280
J Pharmacol Exp Ther. 2002 Nov;303(2):476-86
12388626
Biochem Pharmacol. 2002 Nov 15;64(10):1533-46
12417266
Am J Physiol Renal Physiol. 2003 Apr;284(4):F763-9
12488248
Annu Rev Pharmacol Toxicol. 2004;44:137-66
14744242
Physiol Rev. 2004 Jul;84(3):987-1049
15269342
Nature. 1970 Aug 15;227(5259):680-5
5432063
Biochem J. 1975 Apr;148(1):85-96
1156402
J Surg Res. 1976 Sep;21(3):169-74
957634
FEBS Lett. 1979 Dec 15;108(2):335-40
520571
Proc Natl Acad Sci U S A. 1979 Nov;76(11):5606-10
42902
Biochem Biophys Res Commun. 1980 Sep 30;96(2):848-53
6107079
Biochim Biophys Acta. 1982 Jan 18;700(2):148-53
6120005
FEBS Lett. 1982 Oct 18;147(2):193-6
6129160
Biochem Biophys Res Commun. 1983 Apr 15;112(1):55-60
6838619
J Biol Chem. 1984 Dec 10;259(23):14508-14
6501304
J Biol Chem. 1984 Dec 25;259(24):15393-400
6150935
Annu Rev Pharmacol Toxicol. 1985;25:715-44
3890714
J Biol Chem. 1985 Aug 25;260(18):10293-8
4019514
Am J Physiol. 1985 Aug;249(2 Pt 2):F198-204
2862795
Mol Pharmacol. 1985 Sep;28(3):278-82
3839897
Eur J Biochem. 1986 Jun 16;157(3):605-9
2873036
Kidney Int. 1988 Jul;34(1):74-81
3172638
Pflugers Arch. 1989 Dec;415(3):342-50
2622761
Biochem Pharmacol. 1990 Sep 1;40(5):1131-5
1975172
Toxicol Lett. 1990 Sep;53(1-2):97-104
2219194
J Biol Chem. 1994 Aug 5;269(31):19731-7
8051053
Methods Enzymol. 1995;260:349-69
8592459
Biochem Pharmacol. 1999 Sep 1;58(5):897-907
10449202