title: Characterization of the microsomal steroid-8-ene isomerase of cholesterol biosynthesis.
abstract: Rat liver microsomes contain an enzyme that catalyzes the isomerization of the nuclear double bond of steroids from the 8(9) position to the 7(8) position. The enzyme is most active with zymosterol, 5alpha-cholesta-8,24-dien-3beta-ol, which is a precursor of cholesterol. Properties of the microsomal isomerase have now been studied, and preliminary data are reported on both regulation of enzymic activity and first steps in the solubilization of the enzyme from membranes. After a brief lag period, the velocity of isomerase is relatively constant for about 5 min of incubation, and then isomerization subsides. The apparent Michaelis constant (52-70 micro M) is difficult to determine accurately, due to these complex kinetic changes. V(max) is 4.0-4.7 nmol/min per mg of microsomal protein. The apparent specific activity is more than ten times that of liver microsomal methyl sterol oxidase. The maximal specific activity of microsomal isomerase is approximately doubled when rats are fed an intestinal bile acid sequestrant, cholestyramine. Changes in specific activity of isomerase parallel changes in activities of other microsomal enzymes of cholesterol biosynthesis, such as 3-hydroxy-3-methylglutaryl-CoA reductase and 4-methyl sterol oxidase. Isomerase activity is destroyed by phospholipase A digestion, high concentration of bile salts, and solvent extraction, all of which are known either to remove phospholipid or to alter microsomal membrane integrity. On the other hand, isomerase remains active in the presence of a mild, nonionic detergent, Triton WR-1339; thus, solubilization with nonionic detergents is under study.
mesh_terms: D000818:Animals; D001647:Bile Acids and Salts / Q000494:pharmacology; D002775:Cholestadienols; D002784:Cholesterol / Q000096:biosynthesis*; D002792:Cholestyramine Resin / Q000494:pharmacology; D002793:Cholic Acids / Q000494:pharmacology; D007535:Isomerases / Q000378:metabolism*; D008862:Microsomes, Liver / Q000201:enzymology; D051381:Rats; D013251:Steroid Isomerases / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D001647:Bile Acids and Salts; D002775:Cholestadienols; D002793:Cholic Acids; D002792:Cholestyramine Resin; D002784:Cholesterol; D007535:Isomerases; D013251:Steroid Isomerases; C015582:zymosterol