Functional and Structural Consequences of Nine CYP21A2 Mutations Ranging from Very Mild to Severe Effects
2016 (English)In: International Journal of Endocrinology, ISSN 1687-8337, E-ISSN 1687-8345, 4209670Article in journal (Refereed) Published
Michelatto, Debora de Paula
Univ Estadual Campinas, Ctr Biol Mol & Engn Genet, Lab Genet Mol Humana, Av Candido Rondon 400, BR-13083875 Campinas, SP, Brazil.;Karolinska Univ Hosp, Pediat Endocrinol Unit Q2 08, Karolinska Inst, Dept Womens & Childrens Hlth, S-17176 Stockholm, Sweden..
Karolinska Univ Hosp, Pediat Endocrinol Unit Q2 08, Karolinska Inst, Dept Womens & Childrens Hlth, S-17176 Stockholm, Sweden..
Gori Lusa, Ana Leticia
Univ Estadual Campinas, Ctr Biol Mol & Engn Genet, Lab Genet Mol Humana, Av Candido Rondon 400, BR-13083875 Campinas, SP, Brazil..
Mgnani Silva, Camila D'Almeida
Univ Estadual Campinas, Fac Ciencias Med, Dept Pediat, Rua Tessalia Vieira de Camargo 126, BR-13083887 Campinas, SP, Brazil..
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.
Place, publisher, year, edition, pages
Medical and Health Sciences
IdentifiersURN: urn:nbn:se:uu:diva-307459DOI: 10.1155/2016/4209670ISI: 000385082700001OAI: oai:DiVA.org:uu-307459DiVA: diva2:1047270
FunderMarianne and Marcus Wallenberg FoundationStockholm County CouncilSven Jerring Foundation