Characterization of Organic Anion Transporter 2 (SLC22A7): A Highly Efficient Transporter for Creatinine and Species-Dependent Renal Tubular Expression

Drug Metab Dispos. 2015 Jul;43(7):984-93. doi: 10.1124/dmd.114.062364. Epub 2015 Apr 22.

Abstract

The contribution of organic anion transporter OAT2 (SLC22A7) to the renal tubular secretion of creatinine and its exact localization in the kidney are reportedly controversial. In the present investigation, the transport of creatinine was assessed in human embryonic kidney (HEK) cells that stably expressed human OAT2 (OAT2-HEK) and isolated human renal proximal tubule cells (HRPTCs). The tubular localization of OAT2 in human, monkey, and rat kidney was characterized. The overexpression of OAT2 significantly enhanced the uptake of creatinine in OAT2-HEK cells. Under physiologic conditions (creatinine concentrations of 41.2 and 123.5 µM), the initial rate of OAT2-mediated creatinine transport was approximately 11-, 80-, and 80-fold higher than OCT2, multidrug and toxin extrusion protein (MATE)1, and MATE2K, respectively, resulting in approximately 37-, 1850-, and 80-fold increase of the intrinsic transport clearance when normalized to the transporter protein concentrations. Creatinine intracellular uptake and transcellular transport in HRPTCs were decreased in the presence of 50 µM bromosulfophthalein and 100 µM indomethacin, which inhibited OAT2 more potently than other known creatinine transporters, OCT2 and multidrug and toxin extrusion proteins MATE1 and MATE2K (IC50: 1.3 µM vs. > 100 µM and 2.1 µM vs. > 200 µM for bromosulfophthalein and indomethacin, respectively) Immunohistochemistry analysis showed that OAT2 protein was localized to both basolateral and apical membranes of human and cynomolgus monkey renal proximal tubules, but appeared only on the apical membrane of rat proximal tubules. Collectively, the findings revealed the important role of OAT2 in renal secretion and possible reabsorption of creatinine and suggested a molecular basis for potential species difference in the transporter handling of creatinine.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antiporters / metabolism
  • Creatinine / metabolism*
  • HEK293 Cells
  • Humans
  • Immunohistochemistry
  • Indomethacin / pharmacology
  • Kidney Tubules / metabolism*
  • Kidney Tubules, Proximal / metabolism
  • Kinetics
  • Macaca fascicularis
  • Male
  • Organic Anion Transporters, Sodium-Independent / antagonists & inhibitors
  • Organic Anion Transporters, Sodium-Independent / metabolism*
  • Organic Cation Transport Proteins / metabolism
  • Rats
  • Rats, Sprague-Dawley
  • Species Specificity
  • Sulfobromophthalein / pharmacology

Substances

  • Antiporters
  • Organic Anion Transporters, Sodium-Independent
  • Organic Cation Transport Proteins
  • SLC22A7 protein, human
  • SLC47A1 protein, human
  • SLC47A2 protein, human
  • Slc22a7 protein, rat
  • Slc47a1 protein, rat
  • Sulfobromophthalein
  • Creatinine
  • Indomethacin