Mutants of 11β-hydroxysteroid dehydrogenase (11-HSD2) with partial activity: Improved correlations between genotype and biochemical phenotype in apparent mineralocorticoid excess

B. Scott Nunez, Fraser M. Rogerson, Tomoatsu Mune, Yoshio Igarashi, Yuichi Nakagawa, George Phillipov, Asha Moudgil, Luther B. Travis, Mario Palermo, Cedric Shackleton, Perrin C. White

Research output: Contribution to journalArticle

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Abstract

Mutations in the kidney isozyme of human 11-hydroxysteroid dehydrogenase (11-HSD2) cause apparent mineralocorticoid excess, an autosomal recessive form of familial hypertension. We studied 4 patients with AME, identifying 4 novel and 3 previously reported mutations in the HSD11B2 (HSD11K) gene. Point mutations causing amino acid substitutions were introduced into a pCMV5/11HSD2 expression construct and expressed in mammalian CHOP cells. Mutations L179R and R208H abolished activity in whole cells. Mutants S180F, A237V, and A328V had 19%, 72%, and 25%, respectively, of the activity of the wild-type enzyme in whole cells when cortisol was used as the substrate and 80%, 140%, and 55%, respectively, of wild-type activity when corticosterone was used as the substrate. However, these mutant proteins were only 0.6% to 5.7% as active as the wild-type enzyme in cell lysates, suggesting that these mutations alter stability of the enzyme. In regression analyses of all AME patients with published genotypes, several biochemical and clinical parameters were highly correlated with mutant enzymatic activity, demonstrated in whole cells, when cortisol was used as the substrate. These included the ratio of urinary cortisone to cortisol metabolites (R2=0.648, P<0.0001), age at presentation (R2=0.614, P<0.0001), and birth weight (R2=0.576, P=0.0004). Approximately 5% conversion of cortisol to cortisone is predicted in subjects with mutations that completely inactivate HSD11B2, suggesting that a low level of enzymatic activity is mediated by another enzyme, possibly 11-HSD1.

Original languageEnglish (US)
Pages (from-to)638-642
Number of pages5
JournalHypertension
Volume34
Issue number4 I
StatePublished - Oct 1999

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11-beta-Hydroxysteroid Dehydrogenases
Genotype
Hydrocortisone
Phenotype
Mutation
Cortisone
Enzymes
Enzyme Stability
Mutant Proteins
Amino Acid Substitution
Corticosterone
Point Mutation
Birth Weight
Isoenzymes
Regression Analysis
Apparent mineralocorticoid excess
Hypertension
Kidney
Genes

Keywords

  • Hydrocortisone
  • Hydroxysteroid dehydrogenases
  • Hypertension
  • Metabolism
  • Mineralocortic oids
  • Mutation

ASJC Scopus subject areas

  • Internal Medicine

Cite this

Mutants of 11β-hydroxysteroid dehydrogenase (11-HSD2) with partial activity : Improved correlations between genotype and biochemical phenotype in apparent mineralocorticoid excess. / Nunez, B. Scott; Rogerson, Fraser M.; Mune, Tomoatsu; Igarashi, Yoshio; Nakagawa, Yuichi; Phillipov, George; Moudgil, Asha; Travis, Luther B.; Palermo, Mario; Shackleton, Cedric; White, Perrin C.

In: Hypertension, Vol. 34, No. 4 I, 10.1999, p. 638-642.

Research output: Contribution to journalArticle

Nunez, BS, Rogerson, FM, Mune, T, Igarashi, Y, Nakagawa, Y, Phillipov, G, Moudgil, A, Travis, LB, Palermo, M, Shackleton, C & White, PC 1999, 'Mutants of 11β-hydroxysteroid dehydrogenase (11-HSD2) with partial activity: Improved correlations between genotype and biochemical phenotype in apparent mineralocorticoid excess', Hypertension, vol. 34, no. 4 I, pp. 638-642.
Nunez, B. Scott ; Rogerson, Fraser M. ; Mune, Tomoatsu ; Igarashi, Yoshio ; Nakagawa, Yuichi ; Phillipov, George ; Moudgil, Asha ; Travis, Luther B. ; Palermo, Mario ; Shackleton, Cedric ; White, Perrin C. / Mutants of 11β-hydroxysteroid dehydrogenase (11-HSD2) with partial activity : Improved correlations between genotype and biochemical phenotype in apparent mineralocorticoid excess. In: Hypertension. 1999 ; Vol. 34, No. 4 I. pp. 638-642.
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abstract = "Mutations in the kidney isozyme of human 11-hydroxysteroid dehydrogenase (11-HSD2) cause apparent mineralocorticoid excess, an autosomal recessive form of familial hypertension. We studied 4 patients with AME, identifying 4 novel and 3 previously reported mutations in the HSD11B2 (HSD11K) gene. Point mutations causing amino acid substitutions were introduced into a pCMV5/11HSD2 expression construct and expressed in mammalian CHOP cells. Mutations L179R and R208H abolished activity in whole cells. Mutants S180F, A237V, and A328V had 19{\%}, 72{\%}, and 25{\%}, respectively, of the activity of the wild-type enzyme in whole cells when cortisol was used as the substrate and 80{\%}, 140{\%}, and 55{\%}, respectively, of wild-type activity when corticosterone was used as the substrate. However, these mutant proteins were only 0.6{\%} to 5.7{\%} as active as the wild-type enzyme in cell lysates, suggesting that these mutations alter stability of the enzyme. In regression analyses of all AME patients with published genotypes, several biochemical and clinical parameters were highly correlated with mutant enzymatic activity, demonstrated in whole cells, when cortisol was used as the substrate. These included the ratio of urinary cortisone to cortisol metabolites (R2=0.648, P<0.0001), age at presentation (R2=0.614, P<0.0001), and birth weight (R2=0.576, P=0.0004). Approximately 5{\%} conversion of cortisol to cortisone is predicted in subjects with mutations that completely inactivate HSD11B2, suggesting that a low level of enzymatic activity is mediated by another enzyme, possibly 11-HSD1.",
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T2 - Improved correlations between genotype and biochemical phenotype in apparent mineralocorticoid excess

AU - Nunez, B. Scott

AU - Rogerson, Fraser M.

AU - Mune, Tomoatsu

AU - Igarashi, Yoshio

AU - Nakagawa, Yuichi

AU - Phillipov, George

AU - Moudgil, Asha

AU - Travis, Luther B.

AU - Palermo, Mario

AU - Shackleton, Cedric

AU - White, Perrin C.

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N2 - Mutations in the kidney isozyme of human 11-hydroxysteroid dehydrogenase (11-HSD2) cause apparent mineralocorticoid excess, an autosomal recessive form of familial hypertension. We studied 4 patients with AME, identifying 4 novel and 3 previously reported mutations in the HSD11B2 (HSD11K) gene. Point mutations causing amino acid substitutions were introduced into a pCMV5/11HSD2 expression construct and expressed in mammalian CHOP cells. Mutations L179R and R208H abolished activity in whole cells. Mutants S180F, A237V, and A328V had 19%, 72%, and 25%, respectively, of the activity of the wild-type enzyme in whole cells when cortisol was used as the substrate and 80%, 140%, and 55%, respectively, of wild-type activity when corticosterone was used as the substrate. However, these mutant proteins were only 0.6% to 5.7% as active as the wild-type enzyme in cell lysates, suggesting that these mutations alter stability of the enzyme. In regression analyses of all AME patients with published genotypes, several biochemical and clinical parameters were highly correlated with mutant enzymatic activity, demonstrated in whole cells, when cortisol was used as the substrate. These included the ratio of urinary cortisone to cortisol metabolites (R2=0.648, P<0.0001), age at presentation (R2=0.614, P<0.0001), and birth weight (R2=0.576, P=0.0004). Approximately 5% conversion of cortisol to cortisone is predicted in subjects with mutations that completely inactivate HSD11B2, suggesting that a low level of enzymatic activity is mediated by another enzyme, possibly 11-HSD1.

AB - Mutations in the kidney isozyme of human 11-hydroxysteroid dehydrogenase (11-HSD2) cause apparent mineralocorticoid excess, an autosomal recessive form of familial hypertension. We studied 4 patients with AME, identifying 4 novel and 3 previously reported mutations in the HSD11B2 (HSD11K) gene. Point mutations causing amino acid substitutions were introduced into a pCMV5/11HSD2 expression construct and expressed in mammalian CHOP cells. Mutations L179R and R208H abolished activity in whole cells. Mutants S180F, A237V, and A328V had 19%, 72%, and 25%, respectively, of the activity of the wild-type enzyme in whole cells when cortisol was used as the substrate and 80%, 140%, and 55%, respectively, of wild-type activity when corticosterone was used as the substrate. However, these mutant proteins were only 0.6% to 5.7% as active as the wild-type enzyme in cell lysates, suggesting that these mutations alter stability of the enzyme. In regression analyses of all AME patients with published genotypes, several biochemical and clinical parameters were highly correlated with mutant enzymatic activity, demonstrated in whole cells, when cortisol was used as the substrate. These included the ratio of urinary cortisone to cortisol metabolites (R2=0.648, P<0.0001), age at presentation (R2=0.614, P<0.0001), and birth weight (R2=0.576, P=0.0004). Approximately 5% conversion of cortisol to cortisone is predicted in subjects with mutations that completely inactivate HSD11B2, suggesting that a low level of enzymatic activity is mediated by another enzyme, possibly 11-HSD1.

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KW - Hydroxysteroid dehydrogenases

KW - Hypertension

KW - Metabolism

KW - Mineralocortic oids

KW - Mutation

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