{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"omics_type":["Unknown"],"volume":["40(2)"],"submitter":["Lee GH"],"pubmed_abstract":["Human cytochrome P450 2C19 catalyzes P450 enzyme reactions of various substrates, including steroids and clinical drugs. Recombinant P450 2C19 enzyme with histidine tag was successfully expressed in <i>Escherichia coli</i> and purified using affinity column chromatography. Ultra-performance liquid chromatography-tandem mass (UPLC-MS/MS) spectrometry showed that the purified P450 2C19 enzyme catalyzed 5-hydroxylation reaction of omeprazole. The purified enzyme displayed typical type I binding spectra to progesterone with a <i>K</i><sub>d</sub> value of 4.5 ± 0.2 µM, indicating a tight substrate binding. P450 2C19 catalyzed the hydroxylation of progesterone to produce 21-hydroxy (OH) as a major and 17-OH product as a minor product. Steady-state kinetic analysis of progesterone 21-hydroxylation indicated that the addition of cytochrome <i>b</i><sub>5</sub> stimulated a five-times catalytic turnover number of P450 2C19 with a <i>k</i><sub>cat</sub> value of 1.07 ± 0.08 min<sup>-1</sup>. The molecular docking model of progesterone in the active site of P450 2C19 displayed that the 21-carbon of progesterone was located close to the heme with a distance of 4.7 Å, suggesting 21-hydroxylation of progesterone is the optimal reaction of P450 2C19 enzyme for a productive orientation of the substrate. Our findings will help investigate the extent to which cytochrome <i>b</i><sub>5</sub> affects the metabolism of P450 2C19 to drugs and steroids.<h4>Supplementary information</h4>The online version contains supplementary material available at 10.1007/s43188-023-00219-8."],"journal":["Toxicological research"],"pagination":["215-222"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC10959859"],"repository":["biostudies-literature"],"pubmed_title":["Catalytic enhancements in cytochrome P450 2C19 by cytochrome &lt;i&gt;b&lt;/i&gt;&lt;sub&gt;5&lt;/sub&gt;."],"pmcid":["PMC10959859"],"pubmed_authors":["Kim C","Kim D","Lee YB","Jeong E","Lee SG","Kim V","Lee GH"],"additional_accession":[]},"is_claimable":false,"name":"Catalytic enhancements in cytochrome P450 2C19 by cytochrome &lt;i&gt;b&lt;/i&gt;&lt;sub&gt;5&lt;/sub&gt;.","description":"Human cytochrome P450 2C19 catalyzes P450 enzyme reactions of various substrates, including steroids and clinical drugs. Recombinant P450 2C19 enzyme with histidine tag was successfully expressed in <i>Escherichia coli</i> and purified using affinity column chromatography. Ultra-performance liquid chromatography-tandem mass (UPLC-MS/MS) spectrometry showed that the purified P450 2C19 enzyme catalyzed 5-hydroxylation reaction of omeprazole. The purified enzyme displayed typical type I binding spectra to progesterone with a <i>K</i><sub>d</sub> value of 4.5 ± 0.2 µM, indicating a tight substrate binding. P450 2C19 catalyzed the hydroxylation of progesterone to produce 21-hydroxy (OH) as a major and 17-OH product as a minor product. Steady-state kinetic analysis of progesterone 21-hydroxylation indicated that the addition of cytochrome <i>b</i><sub>5</sub> stimulated a five-times catalytic turnover number of P450 2C19 with a <i>k</i><sub>cat</sub> value of 1.07 ± 0.08 min<sup>-1</sup>. The molecular docking model of progesterone in the active site of P450 2C19 displayed that the 21-carbon of progesterone was located close to the heme with a distance of 4.7 Å, suggesting 21-hydroxylation of progesterone is the optimal reaction of P450 2C19 enzyme for a productive orientation of the substrate. Our findings will help investigate the extent to which cytochrome <i>b</i><sub>5</sub> affects the metabolism of P450 2C19 to drugs and steroids.<h4>Supplementary information</h4>The online version contains supplementary material available at 10.1007/s43188-023-00219-8.","dates":{"release":"2024-01-01T00:00:00Z","publication":"2024 Apr","modification":"2026-06-01T10:13:40.699Z","creation":"2025-07-13T03:04:43.193Z"},"accession":"S-EPMC10959859","cross_references":{"pubmed":["38525137"],"doi":["10.1007/s43188-023-00219-8"]}}