Metabolism of 1α-hydroxyvitamin D3 by cytochrome P450scc to biologically active 1α,20-dihydroxyvitamin D3

Robert C. Tuckey, Zorica Janjetovic, Wei Li, Minh N. Nguyen, Michal A. Zmijewski, Jordan Zjawiony, Andrzej Slominski

Research output: Contribution to journalArticle

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Abstract

Cytochrome P450scc (CYP11A1) metabolizes vitamin D3 to 20-hydroxyvitamin D3 as the major product, with subsequent production of dihydroxy and trihydroxy derivatives. The aim of this study was to determine whether cytochrome P450scc could metabolize 1α-hydroxyvitamin D3 and whether products were biologically active. The major product of 1α-hydroxyvitamin D3 metabolism by P450scc was identified by mass spectrometry and NMR as 1α,20-dihydroxyvitamin D3. Mass spectrometry of minor metabolites revealed the production of another dihydroxyvitamin D3 derivative, two trihydroxy-metabolites made via 1α,20-dihydroxyvitamin D3 and a tetrahydroxyvitamin D3 derivative. The Km for 1α-hydroxyvitamin D3 determined for P450scc incorporated into phospholipid vesicles was 1.4 mol substrate/mol phospholipid, half that observed for vitamin D3. The kcat was 3.0 mol/min/mol P450scc, 6-fold lower than that for vitamin D3. 1α,20-Dihydroxyvitamin D3 inhibited DNA synthesis by human epidermal HaCaT keratinocytes propagated in culture, in a time- and dose-dependent fashion, with a potency similar to that of 1α,25-dihydroxyvitamin D3. 1α,20-Dihydroxyvitamin D3 (10 μM) enhanced CYP24 mRNA levels in HaCaT keratinocytes but the potency was much lower than that reported for 1α,25-dihydroxyvitamin D3. We conclude that the presence of the 1-hydroxyl group in vitamin D3 does not alter the major site of hydroxylation by P450scc which, as for vitamin D3, is at C20. The major product, 1α,20-dihydroxyvitamin D3, displays biological activity on keratinocytes and therefore might be useful pharmacologically.

Original languageEnglish (US)
Pages (from-to)213-219
Number of pages7
JournalJournal of Steroid Biochemistry and Molecular Biology
Volume112
Issue number4-5
DOIs
StatePublished - Dec 1 2008

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Cholesterol Side-Chain Cleavage Enzyme
Metabolism
Cholecalciferol
Keratinocytes
Calcitriol
Metabolites
Derivatives
Mass spectrometry
Mass Spectrometry
Phospholipids
Hydroxylation
Bioactivity
Hydroxyl Radical
dihydroxy-vitamin D3
Nuclear magnetic resonance
Messenger RNA
DNA
Substrates

All Science Journal Classification (ASJC) codes

  • Molecular Medicine
  • Endocrinology, Diabetes and Metabolism
  • Molecular Biology
  • Cell Biology
  • Endocrinology
  • Clinical Biochemistry
  • Biochemistry

Cite this

Metabolism of 1α-hydroxyvitamin D3 by cytochrome P450scc to biologically active 1α,20-dihydroxyvitamin D3. / Tuckey, Robert C.; Janjetovic, Zorica; Li, Wei; Nguyen, Minh N.; Zmijewski, Michal A.; Zjawiony, Jordan; Slominski, Andrzej.

In: Journal of Steroid Biochemistry and Molecular Biology, Vol. 112, No. 4-5, 01.12.2008, p. 213-219.

Research output: Contribution to journalArticle

Tuckey, Robert C. ; Janjetovic, Zorica ; Li, Wei ; Nguyen, Minh N. ; Zmijewski, Michal A. ; Zjawiony, Jordan ; Slominski, Andrzej. / Metabolism of 1α-hydroxyvitamin D3 by cytochrome P450scc to biologically active 1α,20-dihydroxyvitamin D3. In: Journal of Steroid Biochemistry and Molecular Biology. 2008 ; Vol. 112, No. 4-5. pp. 213-219.
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