Dr Hollis served as a consultant for the Diasorin Corporation. All other authors have no conflict of interest.
CYP3A4 is a Human Microsomal Vitamin D 25-Hydroxylase†
Article first published online: 22 DEC 2003
Copyright © 2004 ASBMR
Journal of Bone and Mineral Research
Volume 19, Issue 4, pages 680–688, April 2004
How to Cite
Gupta, R. P., Hollis, B. W., Patel, S. B., Patrick, K. S. and Bell, N. H. (2004), CYP3A4 is a Human Microsomal Vitamin D 25-Hydroxylase. J Bone Miner Res, 19: 680–688. doi: 10.1359/JBMR.0301257
- Issue published online: 2 DEC 2009
- Article first published online: 22 DEC 2003
- Manuscript Accepted: 19 DEC 2003
- Manuscript Revised: 6 NOV 2003
- Manuscript Received: 1 APR 2003
- vitamin D;
- 25-hydroxyvitamin D;
- vitamin D 25-hydroxylase;
- liver microsomes;
- cytochrome P450 enzymes
The human hepatic microsomal vitamin D 25-hydroxylase protein and gene have not been identified with certainty. Sixteen hepatic recombinant microsomal enzymes were screened for 25-hydroxylase activity; 11 had some 25-hydroxylase activity, but CYP3A4 had the highest activity. In characterized liver microsomes, 25-hydroxylase activity correlated significantly with CYP3A4 testosterone 6β-hydroxylase activity. Activity in pooled liver microsomes was inhibited by known inhibitors of CYP3A4 and by an antibody to CYP3A2. Thus, CYP3A4 is a hepatic microsomal vitamin D 25-hydroxylase.
Introduction: Studies were performed to identify human microsomal vitamin D-25 hydroxylase.
Materials and Methods: Sixteen major hepatic microsomal recombinant enzymes derived from cytochrome P450 cDNAs expressed in baculovirus-infected insect cells were screened for 25-hydroxylase activity with 1α-hydroxyvitamin D2 [1α(OH)D2], 1α-hydroxyvitamin D3 [1α(OH)D3], vitamin D2, and vitamin D3 as substrates. Activity was correlated with known biological activities of enzymes in a panel of 12 characterized human liver microsomes. The effects of known inhibitors and specific antibodies on activity also were determined.
Results: CYP3A4, the most abundant cytochrome P450 enzyme in human liver and intestine, had 7-fold greater activity than that of any of the other enzymes with 1α(OH)D2 as substrate. CYP3A4 25-hydroxylase activity was four times higher with 1α(OH)D2 than with 1α(OH)D3 as substrate, was much less with vitamin D2, and was not detected with vitamin D3. 1α(OH)D2 was the substrate in subsequent experiments. In a panel of characterized human liver microsomes, 25-hydroxylase activity correlated with CYP3A4 testosterone 6β-hydroxylase activity (r = 0.93, p < 0.001) and CYP2C91 diclofenac 4′-hydroxylase activity (r = 0.65, p < 0.05), but not with activity of any of the other enzymes. Activity in recombinant CYP3A4 and pooled liver microsomes was dose-dependently inhibited by ketoconazole, troleandomycin, isoniazid, and α-naphthoflavone, known inhibitors of CYP3A4. Activity in pooled liver microsomes was inhibited by antibodies to CYP3A2 that are known to inhibit CYP3A4 activity.
Conclusion: CYP3A4 is a vitamin D 25-hydroxylase for vitamin D2 in human hepatic microsomes and hydroxylates both 1α(OH)D2 and 1α(OH)D3.