{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T18:43:48Z","timestamp":1790793828245,"version":"4.1.0"},"reference-count":30,"publisher":"The Endocrine Society","issue":"7","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2006,7,1]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Context: Congenital adrenal hyperplasia is a group of autosomal recessive inherited disorders of steroidogenesis. The deficiency of steroid 11-hydroxylase (CYP11B1) resulting from mutations in the CYP11B1 gene is the second most frequent cause.<\/jats:p><jats:p>Objective: We studied the functional and structural consequences of two CYP11B1 missense mutations, which were detected in a 1.8-yr-old boy with acne and precocious pseudopuberty, to prove their clinical relevance and study their impact on CYP11B1 function.<\/jats:p><jats:p>Results: The in vitro expression studies in COS-7 cells revealed an almost complete absence of CYP11B1 activity for the P94L mutant to 0.05% for the conversion of 11-deoxycortisol to cortisol. The A368D mutant severely reduced the CYP11B1 enzymatic activity to 1.17%. Intracellular localization studies by immunofluorescence revealed that the mutants were correctly localized. Introducing these mutations in a three-dimensional model structure of the CYP11B1 protein provides a possible explanation for the effects measured in vitro. We hypothesize that the A368D mutation interferes with structures important for substrate specificity and heme iron binding, thus explaining its major functional impact. However, according to structural analysis, we would expect only a minor effect of the P94L mutant on 11-hydroxylase activity, which contrasts with the observed major effect of this mutation both in vitro and in vivo.<\/jats:p><jats:p>Conclusion: Analyzing the in vitro enzyme function is a complementary procedure to genotyping and a valuable tool for understanding the clinical phenotype of 11-hydroxylase deficiency. This is the basis for accurate genetic counseling, prenatal diagnosis, and treatment. Moreover, the combination of in vitro enzyme function and molecular modeling provides valuable insights in cytochrome P450 structural-functional relationships, although one must be aware of the limitations of in silico-based methods.<\/jats:p>","DOI":"10.1210\/jc.2006-0209","type":"journal-article","created":{"date-parts":[[2006,5,3]],"date-time":"2006-05-03T01:29:44Z","timestamp":1146619784000},"page":"2682-2688","source":"Crossref","is-referenced-by-count":36,"title":["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"],"prefix":"10.1210","volume":"91","author":[{"given":"Nils","family":"Krone","sequence":"first","affiliation":[{"name":"Division of Pediatric Endocrinology (N.K., Y.G., M.M., R.E.V., P.-M.H., W.G.S., F.G.R.), Department of Pediatrics, Christian-Albrechts-Universita\u0308t zu Kiel, Universita\u0308tsklinikum Schleswig-Holstein (Campus Kiel), D-24105 Kiel, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yulia","family":"Grischuk","sequence":"additional","affiliation":[{"name":"Division of Pediatric Endocrinology (N.K., Y.G., M.M., R.E.V., P.-M.H., W.G.S., F.G.R.), Department of Pediatrics, Christian-Albrechts-Universita\u0308t zu Kiel, Universita\u0308tsklinikum Schleswig-Holstein (Campus Kiel), D-24105 Kiel, Germany"},{"name":"Engelhardt Institute of Molecular Biology (Y.G.), Russian Academy of Sciences, Moscow 119991, Russian Federation"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marina","family":"Mu\u0308ller","sequence":"additional","affiliation":[{"name":"Division of Pediatric Endocrinology (N.K., Y.G., M.M., R.E.V., P.-M.H., W.G.S., F.G.R.), Department of Pediatrics, Christian-Albrechts-Universita\u0308t zu Kiel, Universita\u0308tsklinikum Schleswig-Holstein (Campus Kiel), D-24105 Kiel, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruth Elisabeth","family":"Volk","sequence":"additional","affiliation":[{"name":"Division of Pediatric Endocrinology (N.K., Y.G., M.M., R.E.V., P.-M.H., W.G.S., F.G.R.), Department of Pediatrics, Christian-Albrechts-Universita\u0308t zu Kiel, Universita\u0308tsklinikum Schleswig-Holstein (Campus Kiel), D-24105 Kiel, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joachim","family":"Gro\u0308tzinger","sequence":"additional","affiliation":[{"name":"Biochemisches Institut (J.G.), Christian-Albrechts-Universita\u0308t zu Kiel, D-24098 Kiel, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paul-Martin","family":"Holterhus","sequence":"additional","affiliation":[{"name":"Division of Pediatric Endocrinology (N.K., Y.G., M.M., R.E.V., P.-M.H., W.G.S., F.G.R.), Department of Pediatrics, Christian-Albrechts-Universita\u0308t zu Kiel, Universita\u0308tsklinikum 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Metab"},{"key":"2020071613543760700_R5","doi-asserted-by":"crossref","first-page":"373","DOI":"10.1002\/humu.1380030408","article-title":"Mutations in steroid 21-hydroxylase (CYP21)","volume":"3","author":"White","year":"1994","journal-title":"Hum Mutat"},{"key":"2020071613543760700_R6","doi-asserted-by":"crossref","first-page":"249","DOI":"10.1055\/s-2002-35389","article-title":"Congenital adrenal hyperplasia: 11\u03b2-hydroxylase deficiency","volume":"20","author":"Peter","year":"2002","journal-title":"Semin Reprod Med"},{"key":"2020071613543760700_R7","first-page":"421","article-title":"Disorders of steroid 11\u03b2-hydroxylase isozymes","volume":"15","author":"White","year":"1994","journal-title":"Endocr Rev"},{"key":"2020071613543760700_R8","first-page":"211","article-title":"Disorders of the aldosterone synthase and steroid 11\u03b2-hydroxylase deficiencies","volume":"51","author":"Peter","year":"1999","journal-title":"Horm 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