title: The human renal 25-hydroxyvitamin D3-1 alpha-hydroxylase: properties studied by isotope-dilution mass spectrometry.
abstract: The renal 25-hydroxyvitamin D3-1 alpha-hydroxylase activity has been measured in normal human kidney cortex, using a highly specific assay based on isotope-dilution mass spectrometry. The cortex was obtained from kidneys removed due to renal tumours. The subcellular distribution of 25-hydroxyvitamin D3-1 alpha-hydroxylase activity was studied. Enzyme activity was only observed in the mitochondrial fraction. Mitochondria from non-tumourous kidney cortex had a Vmax of 0.17 +/- 0.02 pmol min-1 mg-1 protein and the apparent Km was in the range of 14 mumol l-1. There was a tendency to a higher 25-hydroxyvitamin D3-1 alpha-hydroxylase activity in preparations from male kidney (0.21 +/- 0.03 pmol min-1 mg-1 protein) than female (0.12 +/- 0.02, P less than 0.05). A significant inverse correlation between serum phosphate and 25-hydroxyvitamin D3-1 alpha-hydroxylase activity was found. No correlation was observed between enzyme activity and serum levels of 1,25-dihydroxyvitamin D (total and free index), PTH, total calcium or ionized calcium. The results indicate that there is a sex difference in human 25-hydroxyvitamin D3-1 alpha-hydroxylase activity similar to the one observed in laboratory animals. Furthermore, the data support the hypothesis that serum phosphate is a major regulator of 1,25-dihydroxyvitamin D3 production in man.
authors.affiliation: Department of Clinical Chemistry, Karolinska Institute, Huddinge University Hospital, Sweden.
authors.forename: Y
authors.initials: Y
authors.lastname: Hagenfeldt
authors.forename: T
authors.initials: T
authors.lastname: Berlin
mesh_terms: D015090:25-Hydroxyvitamin D3 1-alpha-Hydroxylase / Q000378:metabolism*; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D005260:Female; D006801:Humans; D007672:Kidney Cortex / Q000201:enzymology*; D008297:Male; D013058:Mass Spectrometry / Q000379:methods; D008875:Middle Aged; D008928:Mitochondria / Q000201:enzymology; D010710:Phosphates / Q000097:blood; D012727:Sex Characteristics
chemical_list: D010710:Phosphates; D015090:25-Hydroxyvitamin D3 1-alpha-Hydroxylase