title: Deletion of P399_E401 in NADPH cytochrome P450 oxidoreductase results in partial mixed oxidase deficiency.
abstract: P450 oxidoreductase (POR) is the electron donor for all microsomal P450s including steroidogenic enzymes CYP17A1, CYP19A1 and CYP21A2. We found a novel POR mutation P399_E401del in two unrelated Turkish patients with 46,XX disorder of sexual development. Recombinant POR proteins were produced in yeast and tested for their ability to support steroid metabolizing P450 activities. In comparison to wild-type POR, the P399_E401del protein was found to decrease catalytic efficiency of 21-hydroxylation of progesterone by 68%, 17α-hydroxylation of progesterone by 76%, 17,20-lyase action on 17OH-pregnenolone by 69%, aromatization of androstenedione by 85% and cytochrome c reduction activity by 80%. Protein structure analysis of the three amino acid deletion P399_E401 revealed reduced stability and flexibility of the mutant. In conclusion, P399_E401del is a novel mutation in POR that provides valuable genotype-phenotype and structure-function correlation for mutations in a different region of POR compared to previous studies. Characterization of P399_E401del provides further insight into specificity of different P450s for interaction with POR as well as nature of metabolic disruptions caused by more pronounced effect on specific P450s like CYP17A1 and aromatase.
authors.affiliation: Pediatric Endocrinology, Diabetology and Metabolism, University Children's Hospital, Bern, Switzerland. christa.flueck@dkf.unibe.ch
authors.forename: Christa E
authors.initials: CE
authors.lastname: Flück
authors.forename: Delphine
authors.initials: D
authors.lastname: Mallet
authors.forename: Gaby
authors.initials: G
authors.lastname: Hofer
authors.forename: Dinane
authors.initials: D
authors.lastname: Samara-Boustani
authors.forename: Juliane
authors.initials: J
authors.lastname: Leger
authors.forename: Michel
authors.initials: M
authors.lastname: Polak
authors.forename: Yves
authors.initials: Y
authors.lastname: Morel
authors.forename: Amit V
authors.initials: AV
authors.lastname: Pandey
mesh_terms: D058489:46, XX Disorders of Sex Development / Q000201:enzymology* / Q000235:genetics; D001141:Aromatase / Q000235:genetics / Q000378:metabolism; D002675:Child, Preschool; D005260:Female; D006801:Humans; D007223:Infant; D009251:NADPH-Ferrihemoprotein Reductase / Q000737:chemistry / Q000172:deficiency* / Q000235:genetics; D011487:Protein Conformation; D017384:Sequence Deletion; D013254:Steroid 17-alpha-Hydroxylase / Q000235:genetics / Q000378:metabolism; D013255:Steroid 21-Hydroxylase / Q000235:genetics / Q000378:metabolism
chemical_list: D001141:Aromatase; C561410:CYP19A1 protein, human; C531510:CYP21A2 protein, human; D013255:Steroid 21-Hydroxylase; C501892:CYP17A1 protein, human; D013254:Steroid 17-alpha-Hydroxylase; D009251:NADPH-Ferrihemoprotein Reductase