2. Academic Validation
  3. Functional and Structural Consequences of Nine CYP21A2 Mutations Ranging from Very Mild to Severe Effects

Functional and Structural Consequences of Nine CYP21A2 Mutations Ranging from Very Mild to Severe Effects

  • Int J Endocrinol. 2016;2016:4209670. doi: 10.1155/2016/4209670.
Débora de Paula Michelatto 1,  Leif Karlsson 2,  Ana Letícia Gori Lusa 3,  Camila D'Almeida Mgnani Silva 4,  Linus Joakim Östberg 5,  Bengt Persson 6,  Gil Guerra-Júnior 4,  Sofia Helena Valente de Lemos-Marini 4,  Michela Barbaro 7,  Maricilda Palandi de Mello 3,  Svetlana Lajic 2
Affiliations

Affiliations

  • 1 Laboratório de Genética Molecular Humana, Centro de Biologia Molecular e Engenharia Genética, Universidade Estadual de Campinas, Av. Cândido Rondon 400, 13083-875 Campinas, SP, Brazil; Department of Women's and Children's Health, Karolinska Institutet, Pediatric Endocrinology Unit (Q2:08), Karolinska University Hospital, 171 76 Stockholm, Sweden.
  • 2 Department of Women's and Children's Health, Karolinska Institutet, Pediatric Endocrinology Unit (Q2:08), Karolinska University Hospital, 171 76 Stockholm, Sweden.
  • 3 Laboratório de Genética Molecular Humana, Centro de Biologia Molecular e Engenharia Genética, Universidade Estadual de Campinas, Av. Cândido Rondon 400, 13083-875 Campinas, SP, Brazil.
  • 4 Departamento de Pediatria, Faculdade de Ciências Médicas, Universidade Estadual de Campinas, Rua Tessália Vieira de Camargo 126, 13083-887 Campinas, SP, Brazil.
  • 5 Science for Life Laboratory, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, 171 77 Stockholm, Sweden.
  • 6 Science for Life Laboratory, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, 171 77 Stockholm, Sweden; Science for Life Laboratory, Department of Cell and Molecular Biology, Uppsala University, 751 24 Uppsala, Sweden.
  • 7 Department of Molecular Medicine and Surgery, Karolinska Institutet and Center for Inherited Metabolic Diseases (CMMS L7:05) Karolinska University Hospital, 171 76 Stockholm, Sweden.
PMID: 27721825 DOI: 10.1155/2016/4209670
Abstract

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.

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