Regulation of rat liver microsomal cholesterol 7 alpha-hydroxylase: presence of a cytosolic activator.

Abstract

Microsomal cholesterol 7 alpha-hydroxylase (EC 1.14.13.7) in rat liver was assayed by a single-isotope-incorporation method, and factors influencing its activity were studied. Crude cytosol contained a non-catalytic activator which was heat-stable and on-dialyzable. This activator enhanced cholesterol 7 alpha-hydroxylase catalytic activity. The stimulatory property of this cytosolic activator was not altered by cholestyramine feeding, and was retained after fractionation by ammonium sulfate of saturation up to 65%. 3-Hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) stimulated hydroxylase activity at concentrations up to 90 micro M. ATP at concentration greater than 1.2 mM inhibited hydroxylase activity. NaF was stimulatory at concentrations up to 50 mM with a maximum stimulatory effect at 10 mM, and was antagonistic in effect to ATP. HMG-CoA and ATP at the above inhibitory concentrations and higher abolished the activating effect of the cytosolic factor.