Characterization of Mutations Causing CYP21A2 Deficiency in Brazilian and Portuguese Populations
Deficiency of 21-hydroxylase enzyme (CYP21A2) represents 90% of cases in congenital adrenal hyperplasia (CAH), an autosomal recessive disease caused by defects in cortisol biosynthesis. Computational prediction and functional studies are often the only way to classify variants to understand the link...
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MDPI AG
2021-12-01
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| Series: | International Journal of Molecular Sciences |
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| author | Mayara J. Prado Shripriya Singh Rodrigo Ligabue-Braun Bruna V. Meneghetti Thaiane Rispoli Cristiane Kopacek Karina Monteiro Arnaldo Zaha Maria L. R. Rossetti Amit V. Pandey |
| author_facet | Mayara J. Prado Shripriya Singh Rodrigo Ligabue-Braun Bruna V. Meneghetti Thaiane Rispoli Cristiane Kopacek Karina Monteiro Arnaldo Zaha Maria L. R. Rossetti Amit V. Pandey |
| author_sort | Mayara J. Prado |
| collection | DOAJ |
| description | Deficiency of 21-hydroxylase enzyme (CYP21A2) represents 90% of cases in congenital adrenal hyperplasia (CAH), an autosomal recessive disease caused by defects in cortisol biosynthesis. Computational prediction and functional studies are often the only way to classify variants to understand the links to disease-causing effects. Here we investigated the pathogenicity of uncharacterized variants in the CYP21A2 gene reported in Brazilian and Portuguese populations. Physicochemical alterations, residue conservation, and effect on protein structure were accessed by computational analysis. The enzymatic performance was obtained by functional assay with the wild-type and mutant CYP21A2 proteins expressed in HEK293 cells. Computational analysis showed that p.W202R, p.E352V, and p.R484L have severely impaired the protein structure, while p.P35L, p.L199P, and p.P433L have moderate effects. The p.W202R, p.E352V, p.P433L, and p.R484L variants showed residual 21OH activity consistent with the simple virilizing phenotype. The p.P35L and p.L199P variants showed partial 21OH efficiency associated with the non-classical phenotype. Additionally, p.W202R, p.E352V, and p.R484L also modified the protein expression level. We have determined how the selected CYP21A2 gene mutations affect the 21OH activity through structural and activity alteration contributing to the future diagnosis and management of CYP21A2 deficiency. |
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| language | English |
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| publishDate | 2021-12-01 |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-c922283f80a9499d997022d84cf917622023-11-23T11:37:50ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-12-0123129610.3390/ijms23010296Characterization of Mutations Causing CYP21A2 Deficiency in Brazilian and Portuguese PopulationsMayara J. Prado0Shripriya Singh1Rodrigo Ligabue-Braun2Bruna V. Meneghetti3Thaiane Rispoli4Cristiane Kopacek5Karina Monteiro6Arnaldo Zaha7Maria L. R. Rossetti8Amit V. Pandey9Graduate Program in Cell and Molecular Biology, Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre 91501-970, BrazilDepartment of Biomedical Research, University of Bern, 3010 Bern, SwitzerlandDepartament of Pharmacosciences, Universidade Federal de Ciências da Saúde de Porto Alegre (UFCSPA), Porto Alegre 90050-170, BrazilGraduate Program in Cell and Molecular Biology, Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre 91501-970, BrazilGraduate Program in Cell and Molecular Biology, Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre 91501-970, BrazilServiço de Referência em Triagem Neonatal, Hospital Materno Infantil Presidente Vargas, Porto Alegre 90035-074, BrazilGraduate Program in Cell and Molecular Biology, Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre 91501-970, BrazilGraduate Program in Cell and Molecular Biology, Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre 91501-970, BrazilGraduate Program in Cell and Molecular Biology, Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre 91501-970, BrazilDepartment of Biomedical Research, University of Bern, 3010 Bern, SwitzerlandDeficiency of 21-hydroxylase enzyme (CYP21A2) represents 90% of cases in congenital adrenal hyperplasia (CAH), an autosomal recessive disease caused by defects in cortisol biosynthesis. Computational prediction and functional studies are often the only way to classify variants to understand the links to disease-causing effects. Here we investigated the pathogenicity of uncharacterized variants in the CYP21A2 gene reported in Brazilian and Portuguese populations. Physicochemical alterations, residue conservation, and effect on protein structure were accessed by computational analysis. The enzymatic performance was obtained by functional assay with the wild-type and mutant CYP21A2 proteins expressed in HEK293 cells. Computational analysis showed that p.W202R, p.E352V, and p.R484L have severely impaired the protein structure, while p.P35L, p.L199P, and p.P433L have moderate effects. The p.W202R, p.E352V, p.P433L, and p.R484L variants showed residual 21OH activity consistent with the simple virilizing phenotype. The p.P35L and p.L199P variants showed partial 21OH efficiency associated with the non-classical phenotype. Additionally, p.W202R, p.E352V, and p.R484L also modified the protein expression level. We have determined how the selected CYP21A2 gene mutations affect the 21OH activity through structural and activity alteration contributing to the future diagnosis and management of CYP21A2 deficiency.https://www.mdpi.com/1422-0067/23/1/29621-hydroxylase deficiencycongenital adrenal hyperplasiaCYP21A2functional characterization |
| spellingShingle | Mayara J. Prado Shripriya Singh Rodrigo Ligabue-Braun Bruna V. Meneghetti Thaiane Rispoli Cristiane Kopacek Karina Monteiro Arnaldo Zaha Maria L. R. Rossetti Amit V. Pandey Characterization of Mutations Causing CYP21A2 Deficiency in Brazilian and Portuguese Populations International Journal of Molecular Sciences 21-hydroxylase deficiency congenital adrenal hyperplasia CYP21A2 functional characterization |
| title | Characterization of Mutations Causing CYP21A2 Deficiency in Brazilian and Portuguese Populations |
| title_full | Characterization of Mutations Causing CYP21A2 Deficiency in Brazilian and Portuguese Populations |
| title_fullStr | Characterization of Mutations Causing CYP21A2 Deficiency in Brazilian and Portuguese Populations |
| title_full_unstemmed | Characterization of Mutations Causing CYP21A2 Deficiency in Brazilian and Portuguese Populations |
| title_short | Characterization of Mutations Causing CYP21A2 Deficiency in Brazilian and Portuguese Populations |
| title_sort | characterization of mutations causing cyp21a2 deficiency in brazilian and portuguese populations |
| topic | 21-hydroxylase deficiency congenital adrenal hyperplasia CYP21A2 functional characterization |
| url | https://www.mdpi.com/1422-0067/23/1/296 |
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