Acetoacetyl-CoA synthetase specific activity and concentration in rat tissues.
An antibody against acetoacetyl-CoA synthetase purified from rat liver was raised in rabbits. Utilizing the binding of antibody-antigen complexes to a nitrocellulose membrane, a sensitive enzyme-linked immunosorbent assay was developed to estimate the enzyme concentration in rat tissues. The enzyme concentration (microgram immunoreactive protein/mg protein) in rat liver cytosol was increased about 3-, 1.8- and 7-fold by feeding rats diets containing 5% cholestyramine, 0.2% ML-236B (compactin), and 5% cholestyramine plus 0.2% ML-236B for 4 days, respectively, and decreased about 1.8-fold by fasting the animals or 1.3-fold by feeding them a diet containing 5% cholesterol. Changes in the enzyme activity were almost parallel to those in the enzyme concentration, suggesting the physiological role of this enzyme in cholesterol biosynthesis. Immunoblotting of the hepatic cytosol also confirmed that the increase in enzyme concentration on cholestyramine and/or ML-236B feeding was due to an increase in an enzyme protein the same as the purified enzyme and not the isozymic protein. Among various rat tissues examined, the concentrations of immunologically crossreactive enzyme were higher in lipogenic tissues, such as brain, adipose tissue and liver, than in other tissues. The enzymes in these three tissues were identical in molecular weight determined by gel filtration and immunoblotting.
Ito
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M
Fukui
T
T
Saito
T
T
Tomita
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K
D000818:Animals; D000906:Antibodies; D000924:Anticholesteremic Agents / Q000494:pharmacology; D002791:Cholesterol, Dietary / Q000494:pharmacology; D002792:Cholestyramine Resin / Q000494:pharmacology; D003066:Coenzyme A Ligases / Q000302:isolation & purification / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D005779:Immunodiffusion; D007700:Kinetics; D008099:Liver / Q000187:drug effects / Q000201:enzymology; D008148:Lovastatin / Q000031:analogs & derivatives*; D008297:Male; D009281:Naphthalenes / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D014018:Tissue Distribution
D000906:Antibodies; D000924:Anticholesteremic Agents; D002791:Cholesterol, Dietary; D009281:Naphthalenes; D002792:Cholestyramine Resin; C012258:mevastatin; D008148:Lovastatin; D003066:Coenzyme A Ligases; C023747:acetoacetyl-CoA synthetase