Analyzing the Functional and Structural Consequences of Two Point Mutations (P94L and A368D) in the CYP11B1 Gene Causing Congenital Adrenal Hyperplasia Resulting from 11-Hydroxylase Deficiency

Krone, Nils; Grischuk, Yulia; Müller, Mario; Volk, Ruth; Grötzinger, Joachim; Holterhus, Paul‐Martin; Sippell, Wolfgang G.; Riepe, Felix G.

doi:10.1210/jc.2006-0209

Cited by 41 publications

(33 citation statements)

References 25 publications

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“…We found that 41 compound heterozygotes or homozygotes for select missense or nonsense mutations-namely P49L, R141Q, W260X, G267S, L299P, T318M, T318R, A331V, Q356X, A368D, R374Q, V441G, G444D, G446V, and R448H-present with moderate to severe classic CAH because of 11β-hydroxylase, confirming prior results (3,(14)(15)(16)(17)(22)(23)(24). These patients were mainly from Croatia, Tunisia, Africa, Turkey, and Saudi Arabia.…”

Section: Discussionsupporting

confidence: 88%

Clinical, genetic, and structural basis of congenital adrenal hyperplasia due to 11β-hydroxylase deficiency

Khattab

Haider

Kumar

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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109

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Congenital adrenal hyperplasia (CAH), resulting from mutations in CYP11B1, a gene encoding 11β-hydroxylase, represents a rare autosomal recessive Mendelian disorder of aberrant sex steroid production. Unlike CAH caused by 21-hydroxylase deficiency, the disease is far more common in the Middle East and North Africa, where consanguinity is common often resulting in identical mutations. Clinically, affected female newborns are profoundly virilized (Prader score of 4/5), and both genders display significantly advanced bone ages and are oftentimes hypertensive. We find that 11-deoxycortisol, not frequently measured, is the most robust biochemical marker for diagnosing 11β-hydroxylase deficiency. Finally, computational modeling of 25 missense mutations of CYP11B1 revealed that specific modifications in the heme-binding (R374W and R448C) or substrate-binding (W116C) site of 11β-hydroxylase, or alterations in its stability (L299P and G267S), may predict severe disease. Thus, we report clinical, genetic, hormonal, and structural effects of CYP11B1 gene mutations in the largest international cohort of 108 patients with steroid 11β-hydroxylase deficiency CAH.steroid hormones | missense mutations | classic CAH | ambiguous genitalia C ongenital adrenal hyperplasia (CAH) is a Mendelian disorder transmitted as an autosomal recessive trait. The most prevalent form of CAH arises from steroid 21-hydroxylase enzyme deficiency, accounting for ∼90-95% of all cases (1, 2). In contrast, CAH caused by steroid 11β-hydroxylase deficiency is considerably rare, with a prevalence of 5-8% (3), from which we estimate an overall frequency of 1 in 100,000 live births.Two homologous enzymes, 11β-hydroxylase and aldosterone synthase, are encoded by the CYP11B1 and CYP11B2 genes, respectively. The two genes are 40-kb apart, each comprising nine exons and mapped to chromosome 8q21-22 (3, 4) (Fig. 1A). In contrast to CYP21A2 and its CYP21A1P pseudogene, CYP11B1 and CYP11B2 are both active and do not have a pseudogene. The two encoded homologs, however, have distinct functions in cortisol and aldosterone synthesis, respectively (3). In the zona fasciculata, 11β-hydroxylase converts 11-deoxycortisol and 11-deoxycorticosterone to cortisol and corticosterone, respectively, and is regulated by adrenocorticotropic hormone secreted by the pituitary. In contrast, in the zona glomerulosa aldosterone synthase converts corticosterone to aldosterone with the intermediate production of 18-hydroxycorticosterone. These latter conversions are controlled mainly by the renin angiotensin II system and serum potassium concentration (3).Deficiency of 11β-hydroxylase prevents the conversion of 11-deoxycortisol to cortisol and 11-deoxycorticosterone to corticosterone. This results in high levels of 11-deoxycortisol and 11-deoxycorticosterone, respectively, which are shunted into the androgen synthesis pathway, resulting in high levels of the androgenic steroid, androstenedione. Female newborns are thus profoundly virilized and exhibit significant masculinization of the ex...

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Section: Discussionsupporting

confidence: 88%

Clinical, genetic, and structural basis of congenital adrenal hyperplasia due to 11β-hydroxylase deficiency

Khattab

Haider

Kumar

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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109

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“…Patient 1 is heterozygous for the mutant p.(Arg143Trp), which showed a residual 11b-hydroxylase activity of approximately 8% of wild-type activity, consistent with non-classical 11b-OHD. 12,17,18 The kinetic constants reflect steric changes not necessarily affecting substrate binding but reaction velocity. The residue 143 is located in the C helix with a side chain pointing into the solvent surrounding (Figure 4).…”

Section: Discussionmentioning

confidence: 99%

Congenital adrenal hyperplasia due to 11-beta-hydroxylase deficiency: functional consequences of four CYP11B1 mutations

et al. 2013

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Congenital adrenal hyperplasia (CAH) is one of the most common autosomal recessive inherited endocrine disease. Steroid 11b-hydroxylase deficiency (11b-OHD) is the second most common form of CAH. The aim of the study was to study the functional consequences of three novel and one previously described CYP11B1 gene mutations (p.(Arg143Trp), p.(Ala306Val), p.(Glu310Lys) and p.(Arg332Gln)) detected in patients suffering from classical and non-classical 11b-OHD. Functional analyses were performed by using a HEK293 cell in vitro expression system comparing wild type (WT) with mutant 11b-hydroxylase activity. Mutant proteins were examined in silico to study their effect on the three-dimensional structure of the protein. Two mutations (p.(Ala306Val) and p.(Glu310Lys)) detected in patients with classical 11b-OHD showed a nearly complete loss of 11b-hydroxylase activity. The mutations p.(Arg143Trp) and p.(Arg332Gln) detected in patients with non-classical 11b-OHD showed a partial functional impairment with approximately 8% and 6% of WT activity, respectively. Functional mutation analysis allows the classification of novel CYP11B1 mutations as causes of classical and non-classical 11b-OHD. The detection of patients with non-classical phenotypes underscores the importance to screen patients with a phenotype comparable to non-classical 21-hydroxylase deficiency for mutations in the CYP11B1 gene in case of a negative analysis of the CYP21A2 gene. As CYP11B1 mutations are most often individual for a family, the in vitro analysis of novel mutations is essential for clinical and genetic counselling.

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“…In vitro 11-hydroxylase activity of !5% can be considered severe and is most often associated with classic 11OHD (7,24,40). The patients 1 and 2 carrying a completely inactivating mutation (p.His125Thrfs*8) of the CYP11B1 gene show a typical classical 11OHD phenotype with highly elevated steroids hormones, prenatal virilisation of the external genitalia in females and macrogenitosomia in males (5).…”

Section: Discussionmentioning

confidence: 99%

“…Each mutation was introduced into a CYP11B1-pcDNA3.1 expression vector construct (kindly provided by Prof. R Bernhardt, Institute of Biochemistry, University des Saarlandes, Saarbrücken, Germany) by site-directed mutagenesis as described previously (24,25). In vitro transient transfection assay was performed in triplicates using the HEK293 cell line thrice.…”

Section: Transient Transfection Assaymentioning

confidence: 99%

“…The steroid hormones DOC, B, S and F were simultaneously determined in the cell culture supernatant using an UPLC-MS/MS method as described previously (25,26,27). Western blot analysis was carried out to ensure the expression and translation of WT and mutant proteins as described previously (24,25,28). All assays were performed in at least three independent triplicate experiments, and data are presented as meanGS.D.…”

Section: Transient Transfection Assaymentioning

confidence: 99%

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Characterisation of three novel CYP11B1 mutations in classic and non-classic 11β-hydroxylase deficiency

Polat

Kulle

Karaca

et al. 2014

European Journal of Endocrinology

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Background: Congenital adrenal hyperplasia (CAH) is one of the most common autosomal recessive inherited endocrine diseases. Steroid 11b-hydroxylase (P450c11) deficiency (11OHD) is the second most common form of CAH. Aim: The aim of the study was to study the functional consequences of three novel CYP11B1 gene mutations (p.His125Thrfs*8, p.Leu463_Leu464dup and p.Ser150Leu) detected in patients suffering from 11OHD and to correlate this data with the clinical phenotype. Methods: Functional analyses were done by using a HEK293 cell in vitro expression system comparing WT with mutant P450c11 activity. Mutant proteins were examined in silico to study their effect on the three-dimensional structure of the protein.Results: Two mutations (p.His125Thrfs*8 and p.Leu463_Leu464dup) detected in patients with classic 11OHD showed a complete loss of P450c11 activity. The mutation (p.Ser150Leu) detected in a patient with non-classic 11OHD showed partial functional impairment with 19% of WT activity. Conclusion: Functional mutation analysis enables the correlation of novel CYP11B1 mutations to the classic and non-classic 11OHD phenotype respectively. Mutations causing a non-classic phenotype show typically partial impairment due to reduced maximum reaction velocity comparable with non-classic mutations in 21-hydroxylase deficiency. The increasing number of mutations associated with non-classic 11OHD illustrate that this disease should be considered as diagnosis in patients with otherwise unexplained hyperandrogenism.

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Analyzing the Functional and Structural Consequences of Two Point Mutations (P94L and A368D) in the CYP11B1 Gene Causing Congenital Adrenal Hyperplasia Resulting from 11-Hydroxylase Deficiency

Cited by 41 publications

References 25 publications

Clinical, genetic, and structural basis of congenital adrenal hyperplasia due to 11β-hydroxylase deficiency

Clinical, genetic, and structural basis of congenital adrenal hyperplasia due to 11β-hydroxylase deficiency

Congenital adrenal hyperplasia due to 11-beta-hydroxylase deficiency: functional consequences of four CYP11B1 mutations

Characterisation of three novel CYP11B1 mutations in classic and non-classic 11β-hydroxylase deficiency

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