A new isoform of 11 beta-hydroxysteroid dehydrogenase in aldosterone target cells.
11 beta-Hydroxysteroid dehydrogenase has been proposed to play an important role in aldosterone target cells by degrading endogenous glucocorticoids, thus allowing aldosterone to bind to the relatively nonselective mineralocorticoid receptor. The physiologically important species of this enzyme in renal aldosterone target cells appears to be kinetically and antigenically distinct from the previously characterized liver enzyme. Here we show that 11 beta-steroid dehydrogenase in the microsomal fraction of isolated renal collecting duct cells has a Km for corticosterone of 25.9 +/- 2.4 nM, about 100 times lower than the rat liver enzyme. Surprisingly, the collecting duct enzyme utilizes almost exclusively NAD as cofactor versus NADP used by the liver form. Conversion of corticosterone to 11-dehydrocorticosterone is 2.6 +/- 0.5 and 0.07 +/- 0.01 fmol/min/mg protein with 100 microM of NAD and NADP, respectively, demonstrating a 37.4 +/- 3.5-fold preference for NAD versus NADP. There is practically no conversion of 11-dehydrocorticosterone to corticosterone either with NADH or NADPH, indicating that in collecting duct cells the enzyme operates only in the direction of oxygenation. In addition, 11 beta-steroid dehydrogenase activity is dose dependently inhibited by the end product 11-dehydrocorticosterone while the liver enzyme does not show end product inhibition. We conclude that renal collecting duct cells, the major physiological targets of aldosterone, are protected from circulating glucocorticoids by a hitherto undescribed enzyme of the 11-dehydrogenase family, which differs from the known liver enzyme in having a significantly higher affinity for corticosterone and a different cofactor requirement.
Department of Physiology, Dartmouth Medical School, Lebanon, New Hampshire 03756.
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Rusvai
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Náray-Fejes-Tóth
D042842:11-beta-Hydroxysteroid Dehydrogenases; D000450:Aldosterone / Q000378:metabolism*; D000818:Animals; D006913:Hydroxysteroid Dehydrogenases / Q000302:isolation & purification / Q000378:metabolism*; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism*; D007672:Kidney Cortex / Q000201:enzymology*; D007685:Kidney Tubules, Collecting / Q000201:enzymology*; D007687:Kidney Tubules, Proximal / Q000201:enzymology*; D007700:Kinetics; D011817:Rabbits
D007527:Isoenzymes; D000450:Aldosterone; D006913:Hydroxysteroid Dehydrogenases; D042842:11-beta-Hydroxysteroid Dehydrogenases