title: Partial deficiency of 17α-hydroxylase/17,20-lyase caused by a novel missense mutation in the canonical cytochrome heme-interacting motif.
abstract: BACKGROUND
  Deficiency of 17α-hydroxylase/17,20-lyase is a rare cause of 46,XY disordered sex development.
  
  
  OBJECTIVE
  We characterize in vitro and in vivo effects of two novel CYP17A1 gene mutations identified in a patient with a mild phenotype of CYP17A1 deficiency.
  
  
  METHODS
  A 46,XY patient presented with ambiguous genitalia. CYP17A1 deficiency was suspected at 2 months on the basis of steroid analysis performed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Mutational analysis of the CYP17A1 gene was performed by PCR and Sanger sequencing. To characterize the effect of CYP17A1 mutation on 17α-hydroxylase and 17,20-lyase activities in vitro, HEK293 cells were transiently transfected with CYP17A1 expression plasmids, incubated with progesterone or 17-OH-pregnenolone and concentrations of 17-OH-progesterone or DHEA were then measured in the cell culture medium by LC-MS/MS.
  
  
  RESULTS
  Clinical and hormonal findings in the patient were consistent with partial combined deficiency of 17α-hydroxylase/17,20-lyase. The sequencing of the CYP17A1 gene in the patient revealed compound heterozygosity for two novel mutations: c.107delT p.R36fsX107 and p.W121R. After 6-h in vitro culture of transfected HEK293 cells in the presence of 1 μM progesterone, 17α-hydroxylase activity of p.W121R mutant was 60.5±16.3%, while 17,20-lyase activity of mutant measured from the amount of DHEA produced in the presence of 1 μM of 17-OH-pregnenolone was 15.8±2.6% compared with the WT.
  
  
  CONCLUSIONS
  p.W121R substitution, affecting the first residue in the conserved heme-interacting WXXXR motif of CYP17A1, is associated with partial combined deficiency of 17α-hydroxylase/17,20-lyase.
authors: Department and Laboratory of Inherited Endocrine DisordersEndocrinology Research Centre, Ulitsa Dmitriya Ulianova, 11, Moscow 117036, Russian FederationEngelhardt Institute of Molecular BiologyMoscow 119991, Russian FederationInstitute of Physics and TechnologyMoscow Region 141700, Russian FederationInstitute of General Pathology and PathophysiologyMoscow 125315, Russian Federation Department and Laboratory of Inherited Endocrine DisordersEndocrinology Research Centre, Ulitsa Dmitriya Ulianova, 11, Moscow 117036, Russian FederationEngelhardt Institute of Molecular BiologyMoscow 119991, Russian FederationInstitute of Physics and TechnologyMoscow Region 141700, Russian FederationInstitute of General Pathology and PathophysiologyMoscow 125315, Russian Federation.
authors: Petr
authors: P
authors: Rubtsov
authors: Department and Laboratory of Inherited Endocrine DisordersEndocrinology Research Centre, Ulitsa Dmitriya Ulianova, 11, Moscow 117036, Russian FederationEngelhardt Institute of Molecular BiologyMoscow 119991, Russian FederationInstitute of Physics and TechnologyMoscow Region 141700, Russian FederationInstitute of General Pathology and PathophysiologyMoscow 125315, Russian Federation.
authors: Alexander
authors: A
authors: Nizhnik
authors: Department and Laboratory of Inherited Endocrine DisordersEndocrinology Research Centre, Ulitsa Dmitriya Ulianova, 11, Moscow 117036, Russian FederationEngelhardt Institute of Molecular BiologyMoscow 119991, Russian FederationInstitute of Physics and TechnologyMoscow Region 141700, Russian FederationInstitute of General Pathology and PathophysiologyMoscow 125315, Russian Federation.
authors: Ivan
authors: I
authors: Dedov
authors: Department and Laboratory of Inherited Endocrine DisordersEndocrinology Research Centre, Ulitsa Dmitriya Ulianova, 11, Moscow 117036, Russian FederationEngelhardt Institute of Molecular BiologyMoscow 119991, Russian FederationInstitute of Physics and TechnologyMoscow Region 141700, Russian FederationInstitute of General Pathology and PathophysiologyMoscow 125315, Russian Federation.
authors: Natalia
authors: N
authors: Kalinchenko
authors: Department and Laboratory of Inherited Endocrine DisordersEndocrinology Research Centre, Ulitsa Dmitriya Ulianova, 11, Moscow 117036, Russian FederationEngelhardt Institute of Molecular BiologyMoscow 119991, Russian FederationInstitute of Physics and TechnologyMoscow Region 141700, Russian FederationInstitute of General Pathology and PathophysiologyMoscow 125315, Russian Federation.
authors: Vasily
authors: V
authors: Petrov
authors: Department and Laboratory of Inherited Endocrine DisordersEndocrinology Research Centre, Ulitsa Dmitriya Ulianova, 11, Moscow 117036, Russian FederationEngelhardt Institute of Molecular BiologyMoscow 119991, Russian FederationInstitute of Physics and TechnologyMoscow Region 141700, Russian FederationInstitute of General Pathology and PathophysiologyMoscow 125315, Russian Federation Department and Laboratory of Inherited Endocrine DisordersEndocrinology Research Centre, Ulitsa Dmitriya Ulianova, 11, Moscow 117036, Russian FederationEngelhardt Institute of Molecular BiologyMoscow 119991, Russian FederationInstitute of Physics and TechnologyMoscow Region 141700, Russian FederationInstitute of General Pathology and PathophysiologyMoscow 125315, Russian Federation.
authors: Anna
authors: A
authors: Orekhova
authors: Department and Laboratory of Inherited Endocrine DisordersEndocrinology Research Centre, Ulitsa Dmitriya Ulianova, 11, Moscow 117036, Russian FederationEngelhardt Institute of Molecular BiologyMoscow 119991, Russian FederationInstitute of Physics and TechnologyMoscow Region 141700, Russian FederationInstitute of General Pathology and PathophysiologyMoscow 125315, Russian Federation.
authors: Pavel
authors: P
authors: Spirin
authors: Department and Laboratory of Inherited Endocrine DisordersEndocrinology Research Centre, Ulitsa Dmitriya Ulianova, 11, Moscow 117036, Russian FederationEngelhardt Institute of Molecular BiologyMoscow 119991, Russian FederationInstitute of Physics and TechnologyMoscow Region 141700, Russian FederationInstitute of General Pathology and PathophysiologyMoscow 125315, Russian Federation Department and Laboratory of Inherited Endocrine DisordersEndocrinology Research Centre, Ulitsa Dmitriya Ulianova, 11, Moscow 117036, Russian FederationEngelhardt Institute of Molecular BiologyMoscow 119991, Russian FederationInstitute of Physics and TechnologyMoscow Region 141700, Russian FederationInstitute of General Pathology and PathophysiologyMoscow 125315, Russian Federation.
authors: Vladimir
authors: V
authors: Prassolov
authors: Department and Laboratory of Inherited Endocrine DisordersEndocrinology Research Centre, Ulitsa Dmitriya Ulianova, 11, Moscow 117036, Russian FederationEngelhardt Institute of Molecular BiologyMoscow 119991, Russian FederationInstitute of Physics and TechnologyMoscow Region 141700, Russian FederationInstitute of General Pathology and PathophysiologyMoscow 125315, Russian Federation ant@endocrincentr.ru.
authors: Anatoly
authors: A
authors: Tiulpakov
mesh_terms: D000312:Adrenal Hyperplasia, Congenital / Q000235:genetics*; D000595:Amino Acid Sequence; D003580:Cytochromes / Q000235:genetics*; D004247:DNA / Q000235:genetics; D006418:Heme / Q000235:genetics*; D006801:Humans; D007223:Infant; D008297:Male; D008958:Models, Molecular; D020125:Mutation, Missense / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D013254:Steroid 17-alpha-Hydroxylase / Q000235:genetics / Q000378:metabolism*; D053719:Tandem Mass Spectrometry; D013737:Testis / Q000503:physiopathology
chemical_list: D003580:Cytochromes; D006418:Heme; D004247:DNA; D013254:Steroid 17-alpha-Hydroxylase