Pathways and products for the metabolism of vitamin D3 by cytochrome P450scc

Robert C. Tuckey, Wei Li, Jordan K. Zjawiony, Michal A. Zmijewski, Minh N. Nguyen, Trevor Sweatman, Duane Miller, Andrzej Slominski

Research output: Contribution to journalArticle

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Abstract

Cytochrome P450scc (CYP11A1) can hydroxylate vitamin D3 to produce 20-hydroxyvitamin D3 and other poorly characterized hydroxylated products. The present study aimed to identify all the products of vitamin D3 metabolism by P450scc, as well as the pathways leading to their formation. Besides 20-hydroxyvitamin D3, other major metabolites of vitamin D3 were a dihydroxyvitamin D3 and a trihydroxyvitamin D3 product. The dihydroxyvitamin D3 was clearly identified as 20,23-dihydroxyvitamin D3 by NMR, in contrast to previous reports that postulated hydroxyl groups in positions 20 and 22. NMR of the trihydroxy product identified it as 17α,20,23-trihydroxyvitamin D3. This product could be directly produced by P450scc acting on 20,23-dihydroxyvitamin D3, confirming that hydroxyl groups are present at positions 20 and 23. Three minor products of D3 metabolism by P450scc were identified by MS and by examining their subsequent metabolism by P450scc. These products were 23-hydroxyvitamin D3, 17α-hydroxyvitamin D3 and 17α,20-dihydroxyvitamin D3 and arise from the three P450scc-catalysed hydroxylations occurring in a different order. We conclude that the major pathway of vitamin D3 metabolism by P450scc is: vitamin D3 → 20-hydroxyvitamin D3 → 20,23-dihydroxyvitamin D3 → 17α,20,23-trihydroxyvitamin D3. The major products dissociate from the P450scc active site and accumulate at a concentration well above the P450scc concentration. Our new identification of the major dihydroxyvitamin D3 product as 20,23-dihydroxyvitamin D3, rather than 20,22-dihydroxyvitamin D3, explains why there is no cleavage of the vitamin D3 side chain, unlike the metabolism of cholesterol by P450scc.

Original languageEnglish (US)
Pages (from-to)2585-2596
Number of pages12
JournalFEBS Journal
Volume275
Issue number10
DOIs
StatePublished - May 1 2008

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Cholesterol Side-Chain Cleavage Enzyme
Cholecalciferol
Metabolism
Hydroxyl Radical
Nuclear magnetic resonance
Hydroxylation
Metabolites
Catalytic Domain
Cholesterol
dihydroxy-vitamin D3
20,23-dihydroxyvitamin D3
20-hydroxyvitamin D3

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Tuckey, R. C., Li, W., Zjawiony, J. K., Zmijewski, M. A., Nguyen, M. N., Sweatman, T., ... Slominski, A. (2008). Pathways and products for the metabolism of vitamin D3 by cytochrome P450scc. FEBS Journal, 275(10), 2585-2596. https://doi.org/10.1111/j.1742-4658.2008.06406.x

Pathways and products for the metabolism of vitamin D3 by cytochrome P450scc. / Tuckey, Robert C.; Li, Wei; Zjawiony, Jordan K.; Zmijewski, Michal A.; Nguyen, Minh N.; Sweatman, Trevor; Miller, Duane; Slominski, Andrzej.

In: FEBS Journal, Vol. 275, No. 10, 01.05.2008, p. 2585-2596.

Research output: Contribution to journalArticle

Tuckey, RC, Li, W, Zjawiony, JK, Zmijewski, MA, Nguyen, MN, Sweatman, T, Miller, D & Slominski, A 2008, 'Pathways and products for the metabolism of vitamin D3 by cytochrome P450scc', FEBS Journal, vol. 275, no. 10, pp. 2585-2596. https://doi.org/10.1111/j.1742-4658.2008.06406.x
Tuckey RC, Li W, Zjawiony JK, Zmijewski MA, Nguyen MN, Sweatman T et al. Pathways and products for the metabolism of vitamin D3 by cytochrome P450scc. FEBS Journal. 2008 May 1;275(10):2585-2596. https://doi.org/10.1111/j.1742-4658.2008.06406.x
Tuckey, Robert C. ; Li, Wei ; Zjawiony, Jordan K. ; Zmijewski, Michal A. ; Nguyen, Minh N. ; Sweatman, Trevor ; Miller, Duane ; Slominski, Andrzej. / Pathways and products for the metabolism of vitamin D3 by cytochrome P450scc. In: FEBS Journal. 2008 ; Vol. 275, No. 10. pp. 2585-2596.
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