Purification and regulation of mevalonate kinase from rat liver.
Mevalonate kinase may play a key role in regulating cholesterol biosynthesis because its activity may be regulated via feedback inhibition by intermediates in the cholesterol biosynthetic pathway. To study the regulation of mevalonate kinase, the enzyme was purified to homogeneity from rat liver, and monospecific antibody against mevalonate kinase was prepared. The purified mevalonate kinase had a dimeric structure composed of identical subunits, and the Mr of the enzyme determined by gel chromatography was 86,000. Based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the subunit Mr was 39,900. The pI for mevalonate kinate was 6.2. The levels of mevalonate kinase protein and enzyme activity were determined in the livers of rats treated with either cholesterol-lowering agents (cholestyramine, pravastatin, and lovastatin) or with dietary modifications. Diets containing cholestyramine alone or cholestyramine and either pravastatin or lovastatin increased mevalonate kinase activity 3-6-fold. Mevalonate kinase activity decreased approximately 50% in rats treated with diets containing either 5% cholesterol or 5% cholesterol and 0.5% cholic acid. Fasting did not significantly change mevalonate kinase activity. The amount of mevalonate kinase protein in the liver was quantitated using immunoblots, and the changes in the levels of kinase activity induced by either drug treatment or by cholesterol feeding were correlated with similar changes in the levels of mevalonate kinase protein. Therefore, under these experimental conditions, mevalonate kinase activity in the liver was regulated principally by changes in the rates of enzyme synthesis and degradation.
Tanaka
R D
RD
Department of Cellular Biology, Squibb Institute for Medical Research, Princeton, New Jersey 08543-4000.
Schafer
B L
BL
Lee
L Y
LY
Freudenberger
J S
JS
Mosley
S T
ST
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000924:Anticholesteremic Agents / Q000494:pharmacology; D002792:Cholestyramine Resin / Q000494:pharmacology; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D005260:Female; D005459:Fluorides / Q000494:pharmacology; D006538:Heptanoic Acids / Q000494:pharmacology; D006706:Homeostasis; D006903:Hydroxymethylglutaryl CoA Reductases / Q000378:metabolism; D007700:Kinetics; D008099:Liver / Q000187:drug effects / Q000201:enzymology*; D008148:Lovastatin / Q000494:pharmacology; D008970:Molecular Weight; D009281:Naphthalenes / Q000494:pharmacology; D010770:Phosphotransferases / Q000096:biosynthesis / Q000302:isolation & purification* / Q000378:metabolism; D017853:Phosphotransferases (Alcohol Group Acceptor); D017035:Pravastatin; D051381:Rats; D011919:Rats, Inbred Strains; D012016:Reference Values
D000596:Amino Acids; D000924:Anticholesteremic Agents; D006538:Heptanoic Acids; D009281:Naphthalenes; D002792:Cholestyramine Resin; D008148:Lovastatin; D006903:Hydroxymethylglutaryl CoA Reductases; D010770:Phosphotransferases; D017853:Phosphotransferases (Alcohol Group Acceptor); C021796:mevalonate kinase; D017035:Pravastatin; D005459:Fluorides