{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,4]],"date-time":"2026-08-04T04:03:16Z","timestamp":1785816196549,"version":"3.56.0"},"reference-count":40,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2014,7,7]],"date-time":"2014-07-07T00:00:00Z","timestamp":1404691200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Clinical Endocrinology"],"published-print":{"date-parts":[[2015,1]]},"abstract":"<jats:title>Summary<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>A detailed genotype\u2013phenotype evaluation is presented by studying the enzyme activities of five rare amino acid substitutions (Arg233Gly, Ala265Ser, Arg341Trp, Arg366Cys and Met473Ile) identified in the <jats:italic><jats:styled-content style=\"fixed-case\">CYP<\/jats:styled-content>21A2<\/jats:italic> gene in patients investigated for Congenital adrenal hyperplasia (<jats:styled-content style=\"fixed-case\">CAH<\/jats:styled-content>).<\/jats:p><\/jats:sec><jats:sec><jats:title>Objective<\/jats:title><jats:p>To investigate whether the mutations identified in the <jats:italic><jats:styled-content style=\"fixed-case\">CYP<\/jats:styled-content>21A2<\/jats:italic> gene are disease causing and to establish a gradient for the degree of enzyme impairment to improve prediction of patient phenotype.<\/jats:p><\/jats:sec><jats:sec><jats:title>Design and patients<\/jats:title><jats:p>The <jats:italic><jats:styled-content style=\"fixed-case\">CYP<\/jats:styled-content>21A2<\/jats:italic> genes of seven patients investigated for <jats:styled-content style=\"fixed-case\">CAH<\/jats:styled-content> were sequenced and five mutations were identified. The mutant proteins were expressed <jats:italic>in vitro<\/jats:italic> in <jats:styled-content style=\"fixed-case\">COS<\/jats:styled-content>\u20101 cells, and the enzyme activities towards the two natural substrates were determined to verify the disease\u2010causing state of the mutations. The <jats:italic>in vitro<\/jats:italic> activities of these rare mutations were also compared with the activities of four mutations known to cause nonclassic <jats:styled-content style=\"fixed-case\">CAH<\/jats:styled-content> (Pro30Leu, Val281Leu, Pro453Ser and Pro482Ser) in addition to an <jats:italic>in silico<\/jats:italic> structural evaluation of the novel mutants.<\/jats:p><\/jats:sec><jats:sec><jats:title>Main outcome measure<\/jats:title><jats:p>To verify the disease\u2010causing state of novel mutations.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Five <jats:italic><jats:styled-content style=\"fixed-case\">CYP<\/jats:styled-content>21A2<\/jats:italic> mutations were identified (Arg233Gly, Ala265Ser, Arg341Trp, Arg366Cys and Met473Ile). All mutant proteins exhibited enzyme activities above 5%, and four mutations were classified as nonclassic and one as a normal variant. By comparing the investigated protein changes with four common mutations causing nonclassic <jats:styled-content style=\"fixed-case\">CAH<\/jats:styled-content>, a gradient for the degree of enzyme impairment could be established. Studying rare mutations in <jats:styled-content style=\"fixed-case\">CAH<\/jats:styled-content> increases our knowledge regarding the molecular mechanisms that render a mutation pathogenic. It also improves phenotype predictions and genetic counselling for future generations.<\/jats:p><\/jats:sec>","DOI":"10.1111\/cen.12526","type":"journal-article","created":{"date-parts":[[2014,6,20]],"date-time":"2014-06-20T15:56:29Z","timestamp":1403279789000},"page":"37-44","source":"Crossref","is-referenced-by-count":29,"title":["<i>In vitro<\/i> functional studies of rare <scp>CYP<\/scp>21A2 mutations and establishment of an activity gradient for nonclassic mutations improve phenotype predictions in congenital adrenal hyperplasia"],"prefix":"10.1111","volume":"82","author":[{"given":"Michela","family":"Barbaro","sequence":"first","affiliation":[{"name":"Department of Molecular Medicine and Surgery Karolinska Institutet Stockholm Sweden"},{"name":"Center for Inherited Metabolic Diseases Karolinska University Hospital Stockholm Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fernanda 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