Cortisol to cortisone: glucocorticoid to mineralocorticoid.
11 beta-hydroxysteroid dehydrogenase (11 beta-HSD), by converting cortisol and corticosterone to hormonally inactive cortisone and 11-dehydrocorticosterone, respectively, is an important pre-receptor signaling pathway for the renal mineralocorticoid receptor (MR). This receptor has an equal affinity for the glucocorticoids, cortisol and corticosterone, and for the mineralocorticoid, aldosterone. In states of 11 beta-HSD deficiency such as the syndrome of apparent mineralocorticoid excess (AME) and licorice ingestion, cortisol acts as a potent mineralocorticoid. In addition to the established and cloned type I 11 beta-HSD, a second 11 beta-HSD isoform has been reported in rabbit kidney and human placenta. We have analyzed the kinetics of 11 beta-HSD activity in human kidney and compared it with the expressed human type I 11 beta-HSD cDNA. Microsomes were prepared from mid-gestational human fetal kidneys and incubated with various concentrations of cortisol (0.0125-10 microM) and NAD or NADP. Kinetic analysis revealed a high affinity (apparent Km 60 nM) isoform, the activity of which was exclusively NAD-dependent. No convincing NADP-dependent activity was seen. Similarly with cortisone as a substrate no 11-oxoreductase activity was evident. In contrast, when type I human 11 beta-HSD was ligated into the expression vector pcDNAI and transiently transfected into COS-I cells, low affinity (apparent Km 2.1 microM) NADP-dependent activity was seen. 11-Oxoreductase activity was also observed. The cloned type I human 11 beta-HSD encodes an enzyme with both low-affinity, NADP-dependent, dehydrogenase and 11-oxoreductase activities, but this activity is absent in human fetal kidney (and probably adult kidney).(ABSTRACT TRUNCATED AT 250 WORDS)
Stewart
P M
PM
Department of Medicine, University of Birmingham, Queen Elizabeth Hospital, United Kingdom.
Mason
J I
JI
D042842:11-beta-Hydroxysteroid Dehydrogenases; D000818:Animals; D003348:Cortisone / Q000378:metabolism*; D006801:Humans; D006854:Hydrocortisone / Q000378:metabolism*; D006913:Hydroxysteroid Dehydrogenases / Q000378:metabolism*; D007527:Isoenzymes / Q000378:metabolism*; D007668:Kidney / Q000196:embryology / Q000378:metabolism; D008862:Microsomes, Liver / Q000201:enzymology*; D008901:Mineralocorticoids / Q000378:metabolism*; D051381:Rats; D014407:Tumor Cells, Cultured
D007527:Isoenzymes; D008901:Mineralocorticoids; D006913:Hydroxysteroid Dehydrogenases; D042842:11-beta-Hydroxysteroid Dehydrogenases; D003348:Cortisone; D006854:Hydrocortisone