Busca avançada
Ano de início
Entree


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

Texto completo
Autor(es):
Mostrar menos -
Michelatto, Debora de Paula ; Karlsson, Leif ; Gori Lusa, Ana Leticia ; Mgnani Silva, Camila D'Almeida ; Ostberg, Linus Joakim ; Persson, Bengt ; Guerra-Junior, Gil ; Valente de Lemos-Marini, Sofia Helena ; Barbaro, Michela ; de Mello, Maricilda Palandi ; Lajic, Svetlana
Número total de Autores: 11
Tipo de documento: Artigo Científico
Fonte: INTERNATIONAL JOURNAL OF ENDOCRINOLOGY; v. 2016, p. 10-pg., 2016-01-01.
Resumo

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.Thr450Promutations drastically reduced the enzyme function below 4%. The K-m 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. (AU)

Processo FAPESP: 12/16815-0 - Análise de alterações na expressão gênica e na atividade enzimática resultantes de variações intrônicas e exônicas no gene CYP21A2
Beneficiário:Débora de Paula Michelatto
Modalidade de apoio: Bolsas no Brasil - Doutorado Direto
Processo FAPESP: 14/09844-0 - Análise funcional de novas variações nucleotídicas no gene CYP21A2 identificadas em pacientes com hiperplasia da adrenal congênita
Beneficiário:Débora de Paula Michelatto
Modalidade de apoio: Bolsas no Exterior - Estágio de Pesquisa - Doutorado Direto
Processo FAPESP: 11/51808-2 - Estudo da expressao do gene cyp21a2 e da atividade enzimatica da 21-hidroxilase resultante de mutacoes raras em casos de hyperplasia da adrenal congenita nas formas classica e tardia.
Beneficiário:Maricilda Palandi de Mello
Modalidade de apoio: Auxílio à Pesquisa - Regular