Human cytochrome P450scc (CYP11A1) catalyzes epoxide formation with ergosterol

Robert C. Tuckey, Minh N. Nguyen, Jianjun Chen, Andrzej T. Slominski, Donna M. Baldisseri, Elaine W. Tieu, Jordan K. Zjawiony, Wei Li

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Cytochrome P450scc (P450scc) catalyzes the cleavage of the side chain of both cholesterol and the vitamin D 3 precursor, 7-dehydrocholesterol. The aim of this study was to test the ability of human P450scc to metabolize ergosterol, the vitamin D 2 precursor, and define the structure of the major products. P450scc incorporated into the bilayer of phospholipid vesicles converted ergosterol to two major and four minor products with a k cat of 53 mol · min -1 ·mol P450scc -1 and a K m of 0.18 mol ergosterol/mol phospholipid, similar to the values observed for cholesterol metabolism. The reaction of ergosterol with P450scc was scaled up to make enough of the two major products for structural analysis. From mass spectrometry, NMR, and comparison of the NMR data to that for similar molecules, we determined the structures of the two major products as 20-hydroxy-22,23-epoxy-22,23-dihydroergosterol and 22- keto-23-hydroxy-22,23- dihydroergosterol. Molecular modeling and nuclear Overhauser effect (or enhancement) spectroscopy spectra analysis helped to establish the configurations at C20, C22, and C23 and determine the final structures of major products as 22R,23S-epoxyergosta-5,7-diene-3β,20α-diol and 3β,23S-dihydroxyergosta- 5,7-dien-22-one. It is likely that the formation of the second product is through a 22,23-epoxy (oxirane) intermediate followed by C22 hydroxylation with the formation of strained 22- hydroxy-22,23-epoxide (oxiranol), which is immediately transformed to the more stable α-hydroxyketone. Molecular modeling of ergosterol into the P450scc crystal structure positioned the ergosterol side chain consistent with formation of the above products. Thus, we have shown that P450scc efficiently catalyzes epoxide formation with ergosterol giving rise to novel epoxy, hydroxy, and keto derivatives, without causing cleavage of the side chain.

Original languageEnglish (US)
Pages (from-to)436-444
Number of pages9
JournalDrug Metabolism and Disposition
Volume40
Issue number3
DOIs
StatePublished - Mar 1 2012

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Cholesterol Side-Chain Cleavage Enzyme
Ergosterol
Epoxy Compounds
Phospholipids
Spectrum Analysis
Cholesterol
Ergocalciferols
Ethylene Oxide
Cholecalciferol
Hydroxylation
Mass Spectrometry
Cats

All Science Journal Classification (ASJC) codes

  • Pharmacology
  • Pharmaceutical Science

Cite this

Tuckey, R. C., Nguyen, M. N., Chen, J., Slominski, A. T., Baldisseri, D. M., Tieu, E. W., ... Li, W. (2012). Human cytochrome P450scc (CYP11A1) catalyzes epoxide formation with ergosterol. Drug Metabolism and Disposition, 40(3), 436-444. https://doi.org/10.1124/dmd.111.042515

Human cytochrome P450scc (CYP11A1) catalyzes epoxide formation with ergosterol. / Tuckey, Robert C.; Nguyen, Minh N.; Chen, Jianjun; Slominski, Andrzej T.; Baldisseri, Donna M.; Tieu, Elaine W.; Zjawiony, Jordan K.; Li, Wei.

In: Drug Metabolism and Disposition, Vol. 40, No. 3, 01.03.2012, p. 436-444.

Research output: Contribution to journalArticle

Tuckey, RC, Nguyen, MN, Chen, J, Slominski, AT, Baldisseri, DM, Tieu, EW, Zjawiony, JK & Li, W 2012, 'Human cytochrome P450scc (CYP11A1) catalyzes epoxide formation with ergosterol', Drug Metabolism and Disposition, vol. 40, no. 3, pp. 436-444. https://doi.org/10.1124/dmd.111.042515
Tuckey RC, Nguyen MN, Chen J, Slominski AT, Baldisseri DM, Tieu EW et al. Human cytochrome P450scc (CYP11A1) catalyzes epoxide formation with ergosterol. Drug Metabolism and Disposition. 2012 Mar 1;40(3):436-444. https://doi.org/10.1124/dmd.111.042515
Tuckey, Robert C. ; Nguyen, Minh N. ; Chen, Jianjun ; Slominski, Andrzej T. ; Baldisseri, Donna M. ; Tieu, Elaine W. ; Zjawiony, Jordan K. ; Li, Wei. / Human cytochrome P450scc (CYP11A1) catalyzes epoxide formation with ergosterol. In: Drug Metabolism and Disposition. 2012 ; Vol. 40, No. 3. pp. 436-444.
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abstract = "Cytochrome P450scc (P450scc) catalyzes the cleavage of the side chain of both cholesterol and the vitamin D 3 precursor, 7-dehydrocholesterol. The aim of this study was to test the ability of human P450scc to metabolize ergosterol, the vitamin D 2 precursor, and define the structure of the major products. P450scc incorporated into the bilayer of phospholipid vesicles converted ergosterol to two major and four minor products with a k cat of 53 mol · min -1 ·mol P450scc -1 and a K m of 0.18 mol ergosterol/mol phospholipid, similar to the values observed for cholesterol metabolism. The reaction of ergosterol with P450scc was scaled up to make enough of the two major products for structural analysis. From mass spectrometry, NMR, and comparison of the NMR data to that for similar molecules, we determined the structures of the two major products as 20-hydroxy-22,23-epoxy-22,23-dihydroergosterol and 22- keto-23-hydroxy-22,23- dihydroergosterol. Molecular modeling and nuclear Overhauser effect (or enhancement) spectroscopy spectra analysis helped to establish the configurations at C20, C22, and C23 and determine the final structures of major products as 22R,23S-epoxyergosta-5,7-diene-3β,20α-diol and 3β,23S-dihydroxyergosta- 5,7-dien-22-one. It is likely that the formation of the second product is through a 22,23-epoxy (oxirane) intermediate followed by C22 hydroxylation with the formation of strained 22- hydroxy-22,23-epoxide (oxiranol), which is immediately transformed to the more stable α-hydroxyketone. Molecular modeling of ergosterol into the P450scc crystal structure positioned the ergosterol side chain consistent with formation of the above products. Thus, we have shown that P450scc efficiently catalyzes epoxide formation with ergosterol giving rise to novel epoxy, hydroxy, and keto derivatives, without causing cleavage of the side chain.",
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