Cholesterol biosynthesis from lanosterol: regulation and purification of rat hepatic sterol 8-isomerase.
The membrane bound sterol-8-isomerase (isomerase) catalyzes the anaerobic conversion of sterol-8-ene to the sterol-7-ene isomer in eucaryotes. To examine the regulatory mechanism as well as molecular characteristics of the isomerase we investigated the consequences of alteration of the enzymic activity under various diet conditions. Feeding 5% cholesterol or 0.1% AY-9944 for a minimum of 2 days caused more than a 70% decrease in microsomal isomerase activity. Feeding 5% cholestyramine plus 0.1% lovastatin (CL-diet) for 7 days led to approximately 4.0-fold induction of the isomerase activity. In addition, diurnal variation in the enzymic activity was observed with this diet. Induction of the isomerase activity by the CL-diet was quantitatively reflected in an increase in the cholesterol synthetic rate in isolated rat hepatocytes. The isomerase was highly purified from liver of rats fed the CL-diet, and its molecular mass was determined to be 21,000 Da by denaturing sodium dodecylsulfate gel electrophoresis.
Kang
M K
MK
Department of Biochemistry, Yonsei University, Seoul, Korea.
Kim
C K
CK
Johng
T N
TN
Paik
Y K
YK
D000818:Animals; D000924:Anticholesteremic Agents / Q000494:pharmacology; D002784:Cholesterol / Q000008:administration & dosage / Q000096:biosynthesis* / Q000493:pharmacokinetics; D002940:Circadian Rhythm; D005246:Feedback; D006903:Hydroxymethylglutaryl CoA Reductases / Q000378:metabolism; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; D007700:Kinetics; D007810:Lanosterol / Q000378:metabolism*; D008099:Liver / Q000201:enzymology*; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D013251:Steroid Isomerases / Q000302:isolation & purification* / Q000378:metabolism* / Q000502:physiology; D001371:trans-1,4-Bis(2-chlorobenzaminomethyl)cyclohexane Dihydrochloride / Q000494:pharmacology
D000924:Anticholesteremic Agents; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; D007810:Lanosterol; D001371:trans-1,4-Bis(2-chlorobenzaminomethyl)cyclohexane Dihydrochloride; D002784:Cholesterol; D006903:Hydroxymethylglutaryl CoA Reductases; D013251:Steroid Isomerases; C022669:delta(8)-delta(7)-sterol isomerase