P450 Oxidoreductase Deficiency: Loss of Activity Caused by Protein Instability From a Novel L374H Mutation

Parween, Shaheena; Roucher‐Boulez, Florence; Flück, Christa E.; Lienhardt-Roussie, Anne; Mallet, Delphine; Morel, Yves; Pandey, Amit V.

doi:10.1210/jc.2016-1928

Cited by 32 publications

(43 citation statements)

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“…According to the cases reported in the literature, the most common mutation type in the Japanese population is R457H (51/78, 65.4%), and it is A287P (39/92, 42.4%) in Europeans [6, 11, 13, 16, 25–33], which shows significant ethnic differences. R457H was once thought to be a founder mutation in Japan [34], but this mutation has also been reported in Europeans and other ethnicity [35].…”

Section: Discussionmentioning

confidence: 99%

Cytochrome P450 oxidoreductase deficiency caused by R457H mutation in POR gene in Chinese: case report and literature review

Bai

Wang

2017

J Ovarian Res

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BackgroundCytochrome P450 oxidoreductase deficiency (PORD) is a rare disease exhibiting a variety of clinical manifestations. It can be difficult to differentiate with other diseases such as 21-hydroxylase deficiency (21-OHD), polycystic ovary syndrome (PCOS) and Antley–Bixler syndrome (ABS). Nearly 100 cases of PORD have been reported worldwide. However, the genetic characters and clinical management are still unclear, especially in China.Case presentationIn this study, we report a 27-year-old female Chinese patient who first presented with amenorrhea and recurrence of large ovary cyst. She was misdiagnosed with PCOS and non-classical 21-OHD due to ovary cysts and elevated 17-hydroxy-progesterone. The patient’s complaining of a mild difficulty of bending the metacarpophalangeal joints reminded us to consider PORD, which usually presents with skeletal deformities and sexual dysfunction. The diagnosis of PORD was confirmed by genetic analyses, which showed the patient harboring a homozygous missense mutation in the POR gene (R457H) and her parents carrying the heterozygous mutation. The patient was treated with low-dose corticosteroids and estrogen/progesterone sequential therapy, and her ovarian cyst gradually reduced with regular menstruation in the follow-up. Moreover, the clinical and genetic characteristics of 104 previously reported PORD cases were also summarized and analyzed.ConclusionsPORD is a very rare disease which can be easily misdiagnosed in mild cases. Clinicians should keep in mind of this disease in patients with sexual dysfunction, especially combined with special skeletal deformities. Our data could provide a consciously understanding of this disease for clinic practicers. Low-dose corticosteroids combined with estrogen/progesterone sequential therapy will be effective in PORD patients with recurrence of large ovary cyst. The fact that the reported PORD patients in China carrying an identical variant R457H in POR gene also give us a viewpoint that R457H mutation in POR gene maybe important in causing PORD in Chinese as same as in Japanese.

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Section: Discussionmentioning

confidence: 99%

Cytochrome P450 oxidoreductase deficiency caused by R457H mutation in POR gene in Chinese: case report and literature review

Bai

Wang

2017

J Ovarian Res

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“…The FASTpp assay was performed as described previously [66,79,80]. Reaction mixtures contained 0.2 mg/mL of CYP19A1 WT or polymorphic variants R264H and R264C, 10 mM CaCl2, 0.05 mg/mL thermolysin in 0.1 M potassium phosphate buffer (pH 7.4) containing 20% glycerol, 0.1 % Tween-20, and 1 mM β-mercaptoethanol.…”

Section: Fastpp Assay Of Cyp19a1 and Its Polymorphic Variants R264h Amentioning

confidence: 99%

“…CYP19A1 is a 503 amino acid protein (NP_000094) encoded by the CYP19A1 gene (GeneID:1588, NCBI: NM_000103), located on chromosome 15 (15q21.2, GRCh38 15:51208056-51338597). CYP19A1 is a membrane-bound protein located in the endoplasmic reticulum (ER) and is a member of the may severely affect the production of estrogens, and several variations of POR exist with significant population distribution, which can alter enzymatic activities of CYP19A1 [64][65][66][67][68]. We, therefore, tested the impact of some population variants of POR on estrogen formation by the two variants of CYP19A1.…”

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confidence: 99%

Differential effects of variations in human P450 oxidoreductase on the aromatase activity of CYP19A1 polymorphisms R264C and R264H

Parween

Nardo

Baj

et al. 2019

Preprint

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Aromatase (CYP19A1) converts androgens into estrogens and is required for female sexual development and growth and development in both sexes. CYP19A1 is a member of cytochrome P450 family of heme-thiolate monooxygenases located in the endoplasmic reticulum and depends on reducing equivalents from the reduced nicotinamide adenine dinucleotide phosphate via the cytochrome P450 oxidoreductase coded by POR. Both the CYP19A1 and POR genes are highly polymorphic, and mutations in both these genes are linked to disorders of steroid biosynthesis. We have previously shown that R264C and R264H mutations in CYP19A1, as well as mutations in POR, result in a reduction of CYP19A1 activity. The R264C is a common polymorphic variant of CYP19A1, with high frequency in Asian and African populations. Polymorphic alleles of POR are found in all populations studied so far and, therefore, may influence activities of CYP19A1 allelic variants. So far, effects of variations in POR on enzymatic activities of allelic variants of CYP19A1 or any other steroid metabolizing cytochrome P450 proteins have not been studied. Here we are reporting the effects of three POR variants on the aromatase activities of two CYP19A1 variants, R264C and R264H. We used bacterially expressed and purified preparations of WT and variant forms of CYP19A1 and POR and constructed liposomes with embedded CYP19A1 and POR proteins and assayed the CYP19A1 activities using radiolabeled androstenedione as a substrate. With the WT-POR as a redox partner, the R264C-CYP19A1 showed only 15% of aromatase activity, but the R264H had 87% of aromatase activity compared to WT-CYP19A1. With P284L-POR as a redox partner, R264C-CYP19A1 lost all activity but retained 6.7% of activity when P284T-POR was used as a redox partner. The R264H-CYP19A1 showed low activities with both the POR-P284L as well as the POR-P284T. When the POR-Y607C was used as a redox partner, the R264C-CYP19A1 retained around 5% of CYP19A1 activity. Remarkably, The R264H-CYP19A1 had more than three-fold higher activity compared to WT-CYP19A1 when the POR-Y607C was used as the redox partner, pointing towards a beneficial effect. The slight increase in activity of R264C-CYP19A1 with the P284T-POR and the three-fold increase in activity of the R264H-CYP19A1 with the Y607C-POR point towards a conformational effect and role of protein-protein interaction governed by the R264C and R264H substitutions in the CYP19A1 as well as P284L, P284T and Y607C variants of POR. These studies demonstrate that the allelic variants of P450 when present with a variant form of POR may show different activities, and combined effects of variations in both the P450 enzymes as well as in the POR should be considered when genetic data are available. Recent trends in the whole-exome and whole-genome sequencing as diagnostic tools will permit combined evaluation of variations in multiple genes that are interdependent and may guide treatment options by adjusting therapeutic interventions based on laboratory analysis. Keywords: CYP19A1; Aromatase;...

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“…The protocol for expression of N-27 form of POR variants and subsequent membrane purification is described and adopted from our previous publications Pandey et al, 2007Pandey et al, , 2010Parween et al, 2016;Flück and Pandey, 2017). The cDNAs for WT or mutant POR in pET22b were transformed into Escherichia coli BL21(DE3), single colonies were selected for growth on ampicillin and grown in terrific broth pH 7.4 supplemented with 40 mM FeCl 3 , 4 mM ZnCl 2 , 2 mM CoCl 2 , 2 mM Na 2 MoO 4 , 2 mM CaCl 2 , 2 mM CuCl 2 , 2 mM H 3 BO 3 , 0.5 mg/ml riboflavin, 100 µg/ml carbenicillin at 37 • C to an optical density (OD) 600 nm of 0.6 and temperature was reduced to 25 • C for 16 h. The bacterial cells were collected by centrifugation, washed with PBS and suspended in 100 mM Tris-acetate (pH 7.6), 0.5 M sucrose, and 1 mM EDTA and treated with lysozyme (0.5 mg/ml) and EDTA (0.1 mM [pH 8.0]) at 4 • C for 1 h with slow stirring to generate spheroplasts.…”

Section: Recombinant Expression Of Por and Membrane Purificationmentioning

confidence: 99%

Role of Protein-Protein Interactions in Metabolism: Genetics, Structure, Function

Pandey¹,

Henderson²,

Ishii³

et al. 2018

Frontiers Research Topics

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P450 Oxidoreductase Deficiency: Loss of Activity Caused by Protein Instability From a Novel L374H Mutation

Abstract: This is the first report of a mutation causing PORD by affecting protein stability that causes severe reduction in POR activities. Detailed characterization of individual mutations in POR is required for understanding novel molecular mechanisms causing PORD.

Cited by 32 publications

References 42 publications

Cytochrome P450 oxidoreductase deficiency caused by R457H mutation in POR gene in Chinese: case report and literature review

Cytochrome P450 oxidoreductase deficiency caused by R457H mutation in POR gene in Chinese: case report and literature review

Differential effects of variations in human P450 oxidoreductase on the aromatase activity of CYP19A1 polymorphisms R264C and R264H

Role of Protein-Protein Interactions in Metabolism: Genetics, Structure, Function

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