Purification from pig kidney of a microsomal cytochrome P450 catalyzing 1α-hydroxylation of 25-hydroxyvitamin D3

AbstractA cytochrome P450 catalyzing 1α-hydroxylation of 25-hydroxyvitamin D3 was purified from pig kidney microsomes. The enzyme preparation showed one protein band on gel electrophoresis with apparent Mr of 52,500 and a specific cytochrome P450 content of 10.7 nmol/mg of protein. The 25-hydroxyvitamin D3 1α-hydroxylase copurified with the vitamin D3 25-hydroxylase during purification. A cytochrome P450 catalyzing 1α-hydroxylation was purified also from liver microsomes. The apparently homogeneous enzyme showed the same catalytic properties and apparent Mr as the kidney enzyme. The results of the present communication demonstrate the presence in kidney of a previously unknown microsomal 1α-hydroxylase in addition to the assumed specific mitochondrial 1α-hydroxylase. The possibility that microsomal 1α-hydroxylation in pig kidney and liver is catalyzed by the previously described porcine microsomal vitamin D 25-hydroxylase(s) is discussed