Steroid hydroxylations by human adrenal cortex microsomes 1, 2.

Microsomes were prepared from human adrenals obtained at the time of cadaveric renal transplantation. Microsomes were assayed for cytochrome P-450 concentrations (mean =0.63 nmol/mg protein) and NADPH-cytochrome c reductase activity (mean 65 nmol times min-1 times mg-1 protein). Rates of steroid hydroxylation were measured. In man, the rate of 21-hydroxylation of 17-hydroxyprogesterone was approximately three times the rate of 21-hydroxylation of progesterone. The rate of 17-hydroxylation of progesterone was approximately three times the rate of 21-hydroxylation of progesterone. Substrate binding to microsomes showed a type I spectrum with progesterone and 17-hydroxy-progesterone. Both substrates bound all spectrally identifiable sites. Antibody prepared against procine NADPH-cytochrome c reductase inhibited concomitantly human reductase, 21-hydroxylation of progesterone and 17-hydroxyprogesterone, and 17-hydroxylation of progesterone. These results were compared to previous studies with beef adrenal microsomes. No specific evidence was obtained to suggest multiple forms of 21-hydroxylase in human adrenal microsomes. It appears as though the human adrenal microsomal cytochrome P-450 electron transport chain is immunologically similar to those studied previously--beef adrenal, and rat and human liver.