Effect of Sulfhydryl Modification on Rat Kidney Basolateral Plasma Membrane Transport Function

  • Rais A. Ansari
  • Syed A. A. Rizvi
  • Kazim Husain
  • Anastasios Lymperopoulos
  • William O. Berndt


Transport processes are the hallmark of functioning kidney. Various nephrotoxicants disrupt the transport processes to manifest nephrotoxicity. Of several nephrotoxicants, mercuric chloride (HgCl2) depletes the reduced glutathione (GSH) in kidney and has been observed to affect the in vitro p-aminohippurate (PAH) transport by basolateral (BL) membrane vesicles. The role of renal nonprotein sulfhydryls such as, reduced GSH has been demonstrated to affect the PAH transport by BL membrane vesicles. The role of protein sulfhydryls in transport process of PAH by BL membrane is not known. Due to mercury mediated effects on sulfhydryls, the effects of protein-sulfhydryls (–SH) modifying reagents in the current study were investigated on PAH transport by BL membrane. It was observed that modification of –SH by p-chloromercuribenzoate sulphate (pCMBS), and mercuric chloride (HgCl2) decreased while recovering the protein –SH with dithiothreitol treatment provided protection against the effects of pCMBS, and HgCl2 on PAH transport by BL membrane vesicles.


Transport Membrane function Sulfhydryls Mercuric chloride 


  1. Ansari RA, Thakran RS, Berndt WO (1990) The effects of mercuric chloride on transport by brush border and basolateral membrane vesicles isolated from rat kidney. Toxicol Appl Pharmacol 106:145–153CrossRefGoogle Scholar
  2. Ansari RA, Thakran RS, Berndt WO (1991) Effects of mercuric chloride on renal plasma membrane function after depletion or elevation of renal glutathione. Toxicol Appl Pharmacol 111:364–372CrossRefGoogle Scholar
  3. Astorga B, Wunz TM, Morales M, Wright SH, Pelis RM (2011) Differences in the substrate binding regions of renal organic anion transporters 1 (OAT1) and 3 (OAT3). Am J Physiol Renal Physiol 301:F378–F386CrossRefGoogle Scholar
  4. Baggett JM, Berndt WO (1986) The effect of depletion of nonprotein sulfhydryls by diethyl maleate plus buthionine sulfoximine on renal uptake of mercury in the rat. Toxicol Appl Pharmacol 83:556–562CrossRefGoogle Scholar
  5. Baker MA, Cerniglia GJ, Zaman A (1990) Microtiter plate assay for the measurement of glutathione and glutathione disulfide in large numbers of biological samples. Anal Biochem 190:360–365CrossRefGoogle Scholar
  6. Boumendil-Podevin EF, Podevin RA (1983) Isolation of basolateral and brush-border membranes from the rabbit kidney cortex. Vesicle integrity and membrane sidedness of the basolateral fraction. Biochim Biophys Acta 735:86–94CrossRefGoogle Scholar
  7. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254CrossRefGoogle Scholar
  8. Dantzler WH, Bentley SK (1983) Effects of sulfhydryl reagent, p-chloromercuribenzoate, on p-aminohippurate transport by isolated, perfused snake renal tubules. Renal Physiol 6:209–217Google Scholar
  9. Esteva-Font C, Ballarin J, Fernandez-Llama P (2012) Molecular biology of water and salt regulation in the kidney. Cell Mol Life Sci 69:683–695CrossRefGoogle Scholar
  10. Hori R, Takano M, Okano T, Inui K (1985) Transport of p-aminohippurate, tetraethylammonium and D-glucose in renal brush border membranes from rats with acute renal failure. J Pharmacol Exp Ther 233:776–781Google Scholar
  11. Hosoyamada M, Sekine T, Kanai Y, Endou H (1999) Molecular cloning and functional expression of a multispecific organic anion transporter from human kidney. Am J Physiol 276:F122–F128Google Scholar
  12. Houston MC (2011) Role of mercury toxicity in hypertension, cardiovascular disease, and stroke. J Clin Hypertens (Greenwich) 13:621–627CrossRefGoogle Scholar
  13. Jensen RE, Berndt WO (1988) Epinephrine and norepinephrine enhance p-aminohippurate transport into basolateral membrane vesicles. J Pharmacol Exp Ther 244:543–549Google Scholar
  14. Klonne DR, Johnson DR (1983) Amelioration of mercuric chloride-induced acute renal failure by dithiothreitol. Toxicol Appl Pharmacol 70:459–466CrossRefGoogle Scholar
  15. Lash LH (2011) Renal membrane transport of glutathione in toxicology and disease. Vet Pathol 48:408–419CrossRefGoogle Scholar
  16. Lee HY, Kim KR, Woo JS, Kim YK, Park YS (1990) Transport of organic compounds in renal plasma membrane vesicles of cadmium intoxicated rats. Kidney Int 37:727–735CrossRefGoogle Scholar
  17. Lenaz G (2012) Mitochondria and reactive oxygen species. Which role in physiology and pathology? Adv Exp Med Biol 942:93–136CrossRefGoogle Scholar
  18. McDowell EM, Nagle RB, Zalme RC, McNeil JS, Flamenbaum W, Trump BF (1976) Studies on the pathophysiology of acute renal failure. I. Correlation of ultrastructure and function in the proximal tubule of the rat following administration of mercuric chloride. Virchows Arch B Cell Pathol 22:173–196Google Scholar
  19. Montagna G, Hofer CG, Torres AM (1998) Impairment of cellular redox status and membrane protein activities in kidneys from rats with ischemic acute renal failure. Biochim Biophys Acta 1407:99–108CrossRefGoogle Scholar
  20. Reid G, Wolff NA, Dautzenberg FM, Burckhardt G (1998) Cloning of a human renal p-aminohippurate transporter, hROAT1. Kidney Blood Press Res 21:233–237CrossRefGoogle Scholar
  21. Riedmaier AE, Nies AT, Schaeffeler E, Schwab M (2012) Organic anion transporters and their implications in pharmacotherapy. Pharmacol Rev 64:421–449CrossRefGoogle Scholar
  22. Scatena R (2012) Mitochondria and drugs. Adv Exp Med Biol 942:329–346CrossRefGoogle Scholar
  23. Sekine T, Watanabe N, Hosoyamada M, Kanai Y, Endou H (1997) Expression cloning and characterization of a novel multispecific organic anion transporter. J Biol Chem 272:18526–18529CrossRefGoogle Scholar
  24. Sweet DH, Wolff NA, Pritchard JB (1997) Expression cloning and characterization of ROAT1. The basolateral organic anion transporter in rat kidney. J Biol Chem 272:30088–30095CrossRefGoogle Scholar
  25. Tanaka K, Zhou F, Kuze K, You G (2004) Cysteine residues in the organic anion transporter mOAT1. Biochem J 380:283–287CrossRefGoogle Scholar
  26. Tse SS, Bildstein CL, Liu D, Mamelok RD (1983) Effects of divalent cations and sulfhydryl reagents on the p-aminohippurate (PAH) transporter of renal basal-lateral membranes. J Pharmacol Exp Ther 226:19–26Google Scholar
  27. Zalme RC, McDowell EM, Nagle RB, McNeil JS, Flamenbaum W, Trump BF (1976) Studies on the pathophysiology of acute renal failure. II. A histochemical study of the proximal tubule of the rat following administration of mercuric chloride. Virchows Arch B Cell Pathol 22:197–216Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Rais A. Ansari
    • 1
  • Syed A. A. Rizvi
    • 1
  • Kazim Husain
    • 2
  • Anastasios Lymperopoulos
    • 1
  • William O. Berndt
    • 3
  1. 1.Department of Pharmaceutical Sciences, College of Pharmacy Health Professions DivisionNova Southeastern UniversityFort LauderdaleUSA
  2. 2.Department of Physiology, Pharmacology and ToxicologyPonce School of MedicinePonceUSA
  3. 3.Department of PharmacologyUniversity of Nebraska Medical CenterOmahaUSA

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