Functional and Structural Consequences of Nine CYP21A2 Mutations Ranging from Very Mild to Severe Effects
We present the functional and structural effects of seven novel (p.Leu12Met, p.Arg16Cys, p.Ser101Asn, p.Ser202Gly, p.Pro267Leu, p.Gln389_Ala391del, and p.Thr450Met) and two previously reported but not studied (p.Ser113Phe and p.Thr450Pro) CYP21A2 mutations. Functional analyses were complemented with...
Saved in:
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley
2016-01-01
|
| Series: | International Journal of Endocrinology |
| Online Access: | http://dx.doi.org/10.1155/2016/4209670 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832564850787614720 |
|---|---|
| author | Débora de Paula Michelatto Leif Karlsson Ana Letícia Gori Lusa Camila D’Almeida Mgnani Silva Linus Joakim Östberg Bengt Persson Gil Guerra-Júnior Sofia Helena Valente de Lemos-Marini Michela Barbaro Maricilda Palandi de Mello Svetlana Lajic |
| author_facet | Débora de Paula Michelatto Leif Karlsson Ana Letícia Gori Lusa Camila D’Almeida Mgnani Silva Linus Joakim Östberg Bengt Persson Gil Guerra-Júnior Sofia Helena Valente de Lemos-Marini Michela Barbaro Maricilda Palandi de Mello Svetlana Lajic |
| author_sort | Débora de Paula Michelatto |
| collection | DOAJ |
| description | We present the functional and structural effects of seven novel (p.Leu12Met, p.Arg16Cys, p.Ser101Asn, p.Ser202Gly, p.Pro267Leu, p.Gln389_Ala391del, and p.Thr450Met) and two previously reported but not studied (p.Ser113Phe and p.Thr450Pro) CYP21A2 mutations. Functional analyses were complemented with in silico prediction of mutation pathogenicity based on the recently crystallized human CYP21A2 structure. Mutated proteins were transiently expressed in COS-1 cells and enzyme activities towards 17-hydroxyprogesterone and progesterone were determined. Residual enzyme activities between 43% and 97% were obtained for p.Arg16Cys, p.Ser101Asn, p.Ser202Gly, p.Pro267Leu, and p.Thr450Met, similar to the activities of the well-known nonclassic mutations p.Pro453Ser and p.Pro482Ser, whereas the p.Leu12Met variant showed an activity of 100%. Conversely, the novel p.Ser113Phe, p.Gln389_Ala391del, and p.Thr450Pro mutations drastically reduced the enzyme function below 4%. The Km values for all novel variants were in the same order of magnitude as for the wild-type protein except for p.The450Met. The maximum velocity was decreased for all mutants except for p.Leu12Met. We conclude that p.Leu12Met is a normal variant; the mutations p.Arg16Cys, p.Ser101Asn, p.Ser202Gly, p.Pro267Leu, and p.Thr450Met could be associated with very mild nonclassic CAH, and the mutations p.Ser113Phe, p.Gln389_Ala391del, and p.Thr450Pro are associated with classic CAH. The obtained residual activities indicated a good genotype-phenotype correlation. |
| format | Article |
| id | doaj-art-c14892cd77774b48826f166ccbcde065 |
| institution | Kabale University |
| issn | 1687-8337 1687-8345 |
| language | English |
| publishDate | 2016-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | International Journal of Endocrinology |
| spelling | doaj-art-c14892cd77774b48826f166ccbcde0652025-02-03T01:10:09ZengWileyInternational Journal of Endocrinology1687-83371687-83452016-01-01201610.1155/2016/42096704209670Functional and Structural Consequences of Nine CYP21A2 Mutations Ranging from Very Mild to Severe EffectsDébora de Paula Michelatto0Leif Karlsson1Ana Letícia Gori Lusa2Camila D’Almeida Mgnani Silva3Linus Joakim Östberg4Bengt Persson5Gil Guerra-Júnior6Sofia Helena Valente de Lemos-Marini7Michela Barbaro8Maricilda Palandi de Mello9Svetlana Lajic10Laboratório de Genética Molecular Humana, Centro de Biologia Molecular e Engenharia Genética, Universidade Estadual de Campinas, Av. Cândido Rondon 400, 13083-875 Campinas, SP, BrazilDepartment of Women’s and Children’s Health, Karolinska Institutet, Pediatric Endocrinology Unit (Q2:08), Karolinska University Hospital, 171 76 Stockholm, SwedenLaboratório de Genética Molecular Humana, Centro de Biologia Molecular e Engenharia Genética, Universidade Estadual de Campinas, Av. Cândido Rondon 400, 13083-875 Campinas, SP, BrazilDepartamento de Pediatria, Faculdade de Ciências Médicas, Universidade Estadual de Campinas, Rua Tessália Vieira de Camargo 126, 13083-887 Campinas, SP, BrazilScience for Life Laboratory, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, 171 77 Stockholm, SwedenScience for Life Laboratory, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, 171 77 Stockholm, SwedenDepartamento de Pediatria, Faculdade de Ciências Médicas, Universidade Estadual de Campinas, Rua Tessália Vieira de Camargo 126, 13083-887 Campinas, SP, BrazilDepartamento de Pediatria, Faculdade de Ciências Médicas, Universidade Estadual de Campinas, Rua Tessália Vieira de Camargo 126, 13083-887 Campinas, SP, BrazilDepartment of Molecular Medicine and Surgery, Karolinska Institutet and Center for Inherited Metabolic Diseases (CMMS L7:05) Karolinska University Hospital, 171 76 Stockholm, SwedenLaboratório de Genética Molecular Humana, Centro de Biologia Molecular e Engenharia Genética, Universidade Estadual de Campinas, Av. Cândido Rondon 400, 13083-875 Campinas, SP, BrazilDepartment of Women’s and Children’s Health, Karolinska Institutet, Pediatric Endocrinology Unit (Q2:08), Karolinska University Hospital, 171 76 Stockholm, SwedenWe present the functional and structural effects of seven novel (p.Leu12Met, p.Arg16Cys, p.Ser101Asn, p.Ser202Gly, p.Pro267Leu, p.Gln389_Ala391del, and p.Thr450Met) and two previously reported but not studied (p.Ser113Phe and p.Thr450Pro) CYP21A2 mutations. Functional analyses were complemented with in silico prediction of mutation pathogenicity based on the recently crystallized human CYP21A2 structure. Mutated proteins were transiently expressed in COS-1 cells and enzyme activities towards 17-hydroxyprogesterone and progesterone were determined. Residual enzyme activities between 43% and 97% were obtained for p.Arg16Cys, p.Ser101Asn, p.Ser202Gly, p.Pro267Leu, and p.Thr450Met, similar to the activities of the well-known nonclassic mutations p.Pro453Ser and p.Pro482Ser, whereas the p.Leu12Met variant showed an activity of 100%. Conversely, the novel p.Ser113Phe, p.Gln389_Ala391del, and p.Thr450Pro mutations drastically reduced the enzyme function below 4%. The Km values for all novel variants were in the same order of magnitude as for the wild-type protein except for p.The450Met. The maximum velocity was decreased for all mutants except for p.Leu12Met. We conclude that p.Leu12Met is a normal variant; the mutations p.Arg16Cys, p.Ser101Asn, p.Ser202Gly, p.Pro267Leu, and p.Thr450Met could be associated with very mild nonclassic CAH, and the mutations p.Ser113Phe, p.Gln389_Ala391del, and p.Thr450Pro are associated with classic CAH. The obtained residual activities indicated a good genotype-phenotype correlation.http://dx.doi.org/10.1155/2016/4209670 |
| spellingShingle | Débora de Paula Michelatto Leif Karlsson Ana Letícia Gori Lusa Camila D’Almeida Mgnani Silva Linus Joakim Östberg Bengt Persson Gil Guerra-Júnior Sofia Helena Valente de Lemos-Marini Michela Barbaro Maricilda Palandi de Mello Svetlana Lajic Functional and Structural Consequences of Nine CYP21A2 Mutations Ranging from Very Mild to Severe Effects International Journal of Endocrinology |
| title | Functional and Structural Consequences of Nine CYP21A2 Mutations Ranging from Very Mild to Severe Effects |
| title_full | Functional and Structural Consequences of Nine CYP21A2 Mutations Ranging from Very Mild to Severe Effects |
| title_fullStr | Functional and Structural Consequences of Nine CYP21A2 Mutations Ranging from Very Mild to Severe Effects |
| title_full_unstemmed | Functional and Structural Consequences of Nine CYP21A2 Mutations Ranging from Very Mild to Severe Effects |
| title_short | Functional and Structural Consequences of Nine CYP21A2 Mutations Ranging from Very Mild to Severe Effects |
| title_sort | functional and structural consequences of nine cyp21a2 mutations ranging from very mild to severe effects |
| url | http://dx.doi.org/10.1155/2016/4209670 |
| work_keys_str_mv | AT deboradepaulamichelatto functionalandstructuralconsequencesofninecyp21a2mutationsrangingfromverymildtosevereeffects AT leifkarlsson functionalandstructuralconsequencesofninecyp21a2mutationsrangingfromverymildtosevereeffects AT analeticiagorilusa functionalandstructuralconsequencesofninecyp21a2mutationsrangingfromverymildtosevereeffects AT camiladalmeidamgnanisilva functionalandstructuralconsequencesofninecyp21a2mutationsrangingfromverymildtosevereeffects AT linusjoakimostberg functionalandstructuralconsequencesofninecyp21a2mutationsrangingfromverymildtosevereeffects AT bengtpersson functionalandstructuralconsequencesofninecyp21a2mutationsrangingfromverymildtosevereeffects AT gilguerrajunior functionalandstructuralconsequencesofninecyp21a2mutationsrangingfromverymildtosevereeffects AT sofiahelenavalentedelemosmarini functionalandstructuralconsequencesofninecyp21a2mutationsrangingfromverymildtosevereeffects AT michelabarbaro functionalandstructuralconsequencesofninecyp21a2mutationsrangingfromverymildtosevereeffects AT maricildapalandidemello functionalandstructuralconsequencesofninecyp21a2mutationsrangingfromverymildtosevereeffects AT svetlanalajic functionalandstructuralconsequencesofninecyp21a2mutationsrangingfromverymildtosevereeffects |