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. 1988 Apr;85(7):2311–2314. doi: 10.1073/pnas.85.7.2311

Thiazide diuretic drug receptors in rat kidney: identification with [3H]metolazone.

K Beaumont 1, D A Vaughn 1, D D Fanestil 1
PMCID: PMC279981  PMID: 3353380

Abstract

Thiazides and related diuretics inhibit NaCl reabsorption in the distal tubule through an unknown mechanism. We report here that [3H]metolazone, a diuretic with a thiazide-like mechanism of action, labels a site in rat kidney membranes that has characteristics of the thiazide-sensitive ion transporter. [3H]Metolazone bound with high affinity (Kd = 4.27 nM) to a site with a density of 0.717 pmol/mg of protein in kidney membranes. The binding site was localized to the renal cortex, with little or no binding in other kidney regions and 11 other tissues. The affinities of thiazide-type diuretics for this binding site were significantly correlated with their clinical potency. Halide anions (Cl-, Br-, and I-) specifically inhibited high-affinity binding of [3H]metolazone to this site. [3H]Metolazone also bound with lower affinity (Kd = 289 nM) to sites present in kidney as well as in liver, testis, lung, brain, heart, and other tissues. Calcium antagonists and certain smooth muscle relaxants had Ki values of 0.6-10 microM for these low-affinity sites, which were not inhibited by most of the thiazide diuretics tested. Properties of the high-affinity [3H]metolazone binding site are consistent with its identity as the receptor for thiazide-type diuretics.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BEYER K. H. The mechanism of action of chlorothiazide. Ann N Y Acad Sci. 1958 Feb 3;71(4):363–379. doi: 10.1111/j.1749-6632.1958.tb46763.x. [DOI] [PubMed] [Google Scholar]
  2. Beaumont K., Healy D. P., Fanestil D. D. Autoradiographic localization of benzodiazepine receptors in the rat kidney. Am J Physiol. 1984 Nov;247(5 Pt 2):F718–F724. doi: 10.1152/ajprenal.1984.247.5.F718. [DOI] [PubMed] [Google Scholar]
  3. Bennett W. M., Porter G. A. Efficacy and safety of metolazone in renal failure and the nephrotic syndrome. J Clin Pharmacol. 1973 Aug-Sep;13(8):357–364. doi: 10.1002/j.1552-4604.1973.tb00224.x. [DOI] [PubMed] [Google Scholar]
  4. Brodie R. R., Chasseaud L. F., Walmsley L. M. Determination of the diuretic agent metolazone in plasma by high-performance liquid chromatography. J Chromatogr. 1981 Dec 11;226(2):526–532. doi: 10.1016/s0378-4347(00)86093-4. [DOI] [PubMed] [Google Scholar]
  5. Campbell D. B., Moore R. A. The pharmacology and clinical pharmacology of indapamide. Postgrad Med J. 1981;57 (Suppl 2):7–17. [PubMed] [Google Scholar]
  6. Cheng Y., Prusoff W. H. Relationship between the inhibition constant (K1) and the concentration of inhibitor which causes 50 per cent inhibition (I50) of an enzymatic reaction. Biochem Pharmacol. 1973 Dec 1;22(23):3099–3108. doi: 10.1016/0006-2952(73)90196-2. [DOI] [PubMed] [Google Scholar]
  7. Costanzo L. S., Windhager E. E. Calcium and sodium transport by the distal convoluted tubule of the rat. Am J Physiol. 1978 Nov;235(5):F492–F506. doi: 10.1152/ajprenal.1978.235.5.F492. [DOI] [PubMed] [Google Scholar]
  8. Ellison D. H., Velázquez H., Wright F. S. Thiazide-sensitive sodium chloride cotransport in early distal tubule. Am J Physiol. 1987 Sep;253(3 Pt 2):F546–F554. doi: 10.1152/ajprenal.1987.253.3.F546. [DOI] [PubMed] [Google Scholar]
  9. Eriksson O., Wistrand P. J. A search for a model tissue for studying effects of thiazide diuretics. Acta Physiol Scand. 1987 Feb;129(2):171–179. doi: 10.1111/j.1748-1716.1987.tb08056.x. [DOI] [PubMed] [Google Scholar]
  10. Fernandez P. C., Puschett J. B. Proximal tubular actions of metolazone and chlorothiazide. Am J Physiol. 1973 Oct;225(4):954–961. doi: 10.1152/ajplegacy.1973.225.4.954. [DOI] [PubMed] [Google Scholar]
  11. Kunau R. T., Jr, Weller D. R., Webb H. L. Clarification of the site of action of chlorothiazide in the rat nephron. J Clin Invest. 1975 Aug;56(2):401–407. doi: 10.1172/JCI108105. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lauf P. K., McManus T. J., Haas M., Forbush B., 3rd, Duhm J., Flatman P. W., Saier M. H., Jr, Russell J. M. Physiology and biophysics of chloride and cation cotransport across cell membranes. Fed Proc. 1987 May 15;46(7):2377–2394. [PubMed] [Google Scholar]
  13. Lukeman D. S., Fanestil D. D. Interactions of diuretics with the peripheral-type benzodiazepine receptor in rat kidney. J Pharmacol Exp Ther. 1987 Jun;241(3):950–955. [PubMed] [Google Scholar]
  14. Maren T. H. Current status of membrane-bound carbonic anhydrase. Ann N Y Acad Sci. 1980;341:246–258. doi: 10.1111/j.1749-6632.1980.tb47176.x. [DOI] [PubMed] [Google Scholar]
  15. McPherson G. A. A practical computer-based approach to the analysis of radioligand binding experiments. Comput Programs Biomed. 1983 Aug-Oct;17(1-2):107–113. doi: 10.1016/0010-468x(83)90031-4. [DOI] [PubMed] [Google Scholar]
  16. Munson P. J., Rodbard D. Ligand: a versatile computerized approach for characterization of ligand-binding systems. Anal Biochem. 1980 Sep 1;107(1):220–239. doi: 10.1016/0003-2697(80)90515-1. [DOI] [PubMed] [Google Scholar]
  17. Saier M. H., Jr, Boyden D. A. Mechanism, regulation and physiological significance of the loop diuretic-sensitive NaCl/KCl symport system in animal cells. Mol Cell Biochem. 1984;59(1-2):11–32. doi: 10.1007/BF00231303. [DOI] [PubMed] [Google Scholar]
  18. Shetty B. V., Campanella L. A., Thomas T. L., Fedorchuk M., Davidson T. A., Michelson L., Volz H., Zimmerman S. E. Synthesis and activity of some 3-aryl- and 3-aralkyl-1,2,3,4-tetrahydro-4-oxo-6-quinazolinesulfonamides. J Med Chem. 1970 Sep;13(5):886–895. doi: 10.1021/jm00299a022. [DOI] [PubMed] [Google Scholar]
  19. Stokes J. B. Sodium chloride absorption by the urinary bladder of the winter flounder. A thiazide-sensitive, electrically neutral transport system. J Clin Invest. 1984 Jul;74(1):7–16. doi: 10.1172/JCI111420. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Wistrand P. J. Properties of membrane-bound carbonic anhydrase. Ann N Y Acad Sci. 1984;429:195–206. doi: 10.1111/j.1749-6632.1984.tb12333.x. [DOI] [PubMed] [Google Scholar]

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