The steroid biosynthetic enzymes in the adrenal cortex are localised in endoplasmic reticulum and mitochondrial membranes. For some of the enzymes in endoplasmic reticulum the activity appears to be modulated by lipid fluidity, (21-hydroxysteroid hydroxylase and 3 beta-hydroxysteroid dehydrogenase). A mechanism for the regulation of corticosteroid biosynthesis mediated by the membrane fluidity has been suggested. Therefore a study of the mitochondrial inner membrane of the bovine adrenal cortex has been undertaken in comparison with a previous study of the endoplasmic reticulum. The kinetic parameters of the 3 beta-hydroxysteroid dehydrogenase were studied as a function of pH and temperature. No thermal transition can be observed in the Arrhenius plot for this enzyme in contrast with the results obtained for the microsomal enzyme. Membrane fluidity using, as fluorescent probes, diphenylhexatriene and a set of n-(9-anthroyloxy)fatty acids has been also studied as a function of temperature with or without addition of cholesterol. No thermal transition in the lipid phase can be observed. The addition of cholesterol to total mitochondrial membrane as to a lipid extract of the membrane decreases fluidity to the same extent as it does with microsomes. The presence of a large amount of protein in mitochondria has an effect which is additive to that of the cholesterol.
3-Hydroxysteroid Dehydrogenases Adrenal Cortex