Identification of 25,26-dihydroxyvitamin D3 as a rat renal 25-hydroxyvitamin D3 metabolite

25,26-Dihydroxyvitamin D3 [25,26-(OH)2D3] was unequivocally identified as a major renal microsomal metabolite of 25-hydroxyvitamin D3 in rats fed a vitamin D sufficient diet. The structural assignment was based on a comparison of the high-performance liquid chromatograms of synthetic and in vitro ge...

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Bibliographic Details
Published in:Biochemistry (Easton) 1981-09, Vol.20 (20), p.5865-5871
Main Authors: Napoli, Joseph L, Okita, Richard T, Masters, Bettie Sue, Horst, Ronald L
Format: Article
Language:English
Subjects:
Animals
Calcifediol
Chromatography, High Pressure Liquid
Dihydroxycholecalciferols - metabolism
Hydroxycholecalciferols - analysis
Hydroxycholecalciferols - metabolism
Kidney - metabolism
Male
Mass Spectrometry
Microsomes - analysis
Rats
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Summary:25,26-Dihydroxyvitamin D3 [25,26-(OH)2D3] was unequivocally identified as a major renal microsomal metabolite of 25-hydroxyvitamin D3 in rats fed a vitamin D sufficient diet. The structural assignment was based on a comparison of the high-performance liquid chromatograms of synthetic and in vitro generated 25,26-(OH)2D3 through four different systems, the ultraviolet absorbance and mass spectral characteristics of biological 25,26-(OH)2D3, and the chromatographic and mass spectral characteristics of the sodium metaperiodate cleavage product of the metabolite. The enzymic synthesis of 25,26-(OH)2D3 was inhibited 60--80% by a semipurified goat anti-rat NADPH--cytochrome P-450 reductase. This implicates cytochrome P-450 as the probable terminal oxidase of the 25-hydroxyvitamin D3-26-hydroxylase system. The methodology used to assay rat renal 25-OH-D3-hydroxylases is also discussed.
ISSN:0006-2960
1520-4995
DOI:10.1021/bi00523a033