In a recent work we reported that a preparation of cytochrome P-450 from rat liver mitochondria was able to catalyze 25-hydroxylation of vitamin DI when combined with NADPH, adrenal ferredoxin, and ferredoxin reductase (Pedersen, J.-I., Holmberg, I., and Bjorkhem, I. (1979) FEBS Lett.98, 394).Optimal assay conditions have now been worked out for this conversion using a method based on isotope dilution-mass fragmentography.It was shown that a l l three components of the reconstituted system were required for the hydroxylation activity.The rate of conversion (about 0.02 nmol/nmol of cytochrome P-
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Björkhem et al. (1980) studied this question.
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