Exploring Novel Variants of the Cytochrome P450 Reductase Gene (<i>POR</i>) from the Genome Aggregation Database by Integrating Bioinformatic Tools and Functional Assays
Cytochrome P450 oxidoreductase (POR) is an essential redox partner for steroid and drug-metabolizing cytochromes P450 located in the endoplasmic reticulum. Mutations in <i>POR</i> lead to metabolic disorders, including congenital adrenal hyperplasia, and affect the metabolism of steroids...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-11-01
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| Series: | Biomolecules |
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| Online Access: | https://www.mdpi.com/2218-273X/13/12/1728 |
| _version_ | 1797381898907942912 |
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| author | Maria Natalia Rojas Velazquez Søren Therkelsen Amit V. Pandey |
| author_facet | Maria Natalia Rojas Velazquez Søren Therkelsen Amit V. Pandey |
| author_sort | Maria Natalia Rojas Velazquez |
| collection | DOAJ |
| description | Cytochrome P450 oxidoreductase (POR) is an essential redox partner for steroid and drug-metabolizing cytochromes P450 located in the endoplasmic reticulum. Mutations in <i>POR</i> lead to metabolic disorders, including congenital adrenal hyperplasia, and affect the metabolism of steroids, drugs, and xenobiotics. In this study, we examined approximately 450 missense variants of the <i>POR</i> gene listed in the Genome Aggregation Database (gnomAD) using eleven different in silico prediction tools. We found that 64 novel variants were consistently predicted to be disease-causing by most tools. To validate our findings, we conducted a population analysis and selected two variations in <i>POR</i> for further investigation. The human POR wild type and the R268W and L577P variants were expressed in bacteria and subjected to enzyme kinetic assays using a model substrate. We also examined the activities of several cytochrome P450 proteins in the presence of POR (WT or variants) by combining P450 and reductase proteins in liposomes. We observed a decrease in enzymatic activities (ranging from 35% to 85%) of key drug-metabolizing enzymes, supported by POR variants R288W and L577P compared to WT-POR. These results validate our approach of curating a vast amount of data from genome projects and provide an updated and reliable reference for diagnosing POR deficiency. |
| first_indexed | 2024-03-08T20:58:09Z |
| format | Article |
| id | doaj.art-d858fead6721452aacf50dd7e6ad4754 |
| institution | Directory Open Access Journal |
| issn | 2218-273X |
| language | English |
| last_indexed | 2024-03-08T20:58:09Z |
| publishDate | 2023-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biomolecules |
| spelling | doaj.art-d858fead6721452aacf50dd7e6ad47542023-12-22T13:55:55ZengMDPI AGBiomolecules2218-273X2023-11-011312172810.3390/biom13121728Exploring Novel Variants of the Cytochrome P450 Reductase Gene (<i>POR</i>) from the Genome Aggregation Database by Integrating Bioinformatic Tools and Functional AssaysMaria Natalia Rojas Velazquez0Søren Therkelsen1Amit V. Pandey2Division of Pediatric Endocrinology, Department of Pediatrics, University Children’s Hospital Bern, 3010 Bern, SwitzerlandDivision of Pediatric Endocrinology, Department of Pediatrics, University Children’s Hospital Bern, 3010 Bern, SwitzerlandDivision of Pediatric Endocrinology, Department of Pediatrics, University Children’s Hospital Bern, 3010 Bern, SwitzerlandCytochrome P450 oxidoreductase (POR) is an essential redox partner for steroid and drug-metabolizing cytochromes P450 located in the endoplasmic reticulum. Mutations in <i>POR</i> lead to metabolic disorders, including congenital adrenal hyperplasia, and affect the metabolism of steroids, drugs, and xenobiotics. In this study, we examined approximately 450 missense variants of the <i>POR</i> gene listed in the Genome Aggregation Database (gnomAD) using eleven different in silico prediction tools. We found that 64 novel variants were consistently predicted to be disease-causing by most tools. To validate our findings, we conducted a population analysis and selected two variations in <i>POR</i> for further investigation. The human POR wild type and the R268W and L577P variants were expressed in bacteria and subjected to enzyme kinetic assays using a model substrate. We also examined the activities of several cytochrome P450 proteins in the presence of POR (WT or variants) by combining P450 and reductase proteins in liposomes. We observed a decrease in enzymatic activities (ranging from 35% to 85%) of key drug-metabolizing enzymes, supported by POR variants R288W and L577P compared to WT-POR. These results validate our approach of curating a vast amount of data from genome projects and provide an updated and reliable reference for diagnosing POR deficiency.https://www.mdpi.com/2218-273X/13/12/1728cytochrome P450 oxidoreductasePORgnomADCAHdrug metabolismsteroid hormones |
| spellingShingle | Maria Natalia Rojas Velazquez Søren Therkelsen Amit V. Pandey Exploring Novel Variants of the Cytochrome P450 Reductase Gene (<i>POR</i>) from the Genome Aggregation Database by Integrating Bioinformatic Tools and Functional Assays Biomolecules cytochrome P450 oxidoreductase POR gnomAD CAH drug metabolism steroid hormones |
| title | Exploring Novel Variants of the Cytochrome P450 Reductase Gene (<i>POR</i>) from the Genome Aggregation Database by Integrating Bioinformatic Tools and Functional Assays |
| title_full | Exploring Novel Variants of the Cytochrome P450 Reductase Gene (<i>POR</i>) from the Genome Aggregation Database by Integrating Bioinformatic Tools and Functional Assays |
| title_fullStr | Exploring Novel Variants of the Cytochrome P450 Reductase Gene (<i>POR</i>) from the Genome Aggregation Database by Integrating Bioinformatic Tools and Functional Assays |
| title_full_unstemmed | Exploring Novel Variants of the Cytochrome P450 Reductase Gene (<i>POR</i>) from the Genome Aggregation Database by Integrating Bioinformatic Tools and Functional Assays |
| title_short | Exploring Novel Variants of the Cytochrome P450 Reductase Gene (<i>POR</i>) from the Genome Aggregation Database by Integrating Bioinformatic Tools and Functional Assays |
| title_sort | exploring novel variants of the cytochrome p450 reductase gene i por i from the genome aggregation database by integrating bioinformatic tools and functional assays |
| topic | cytochrome P450 oxidoreductase POR gnomAD CAH drug metabolism steroid hormones |
| url | https://www.mdpi.com/2218-273X/13/12/1728 |
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