Biochemical evidence for oligomerization of rat adrenal acyl-coenzyme A:cholesterol acyltransferase. 1998

T Kawasaki, and A Miyazaki, and H Hakamata, and H Matsuda, and S Horiuchi
Department of Biochemistry, Kumamoto University School of Medicine, Japan.

Acyl-coenzyme A:cholesterol acyltransferase (ACAT) in rat adrenal was compared with that in rat liver. Immunoblot analyses of the microsomal fractions from adrenal with an anti-human ACAT antibody detected a 45 kDa protein. Upon pretreatment of these microsomal fractions with chemical cross-linkers such as BS3 and Sulfo-EGS, the 45 kDa band decreased with a concomitant increase in high molecular weight proteins (55, approximately 100, and approximately 230 kDa), suggesting that ACAT constitutes oligomers of 45 kDa monomers associated with a 10 kDa protein. In sharp contrast, the same immunoblot analysis of rat liver microsomal fractions identified a 50 kDa protein which was not cross-linked by these cross-linkers. Moreover, when four ACAT inhibitors were tested for their effects on adrenal and liver enzymes, NTE-122, CI-976, and E5324 were more effective for the liver enzyme, whereas 58-035 was much more effective for adrenal ACAT. These biochemical and pharmacological observations support the notion that the rat liver ACAT protein is distinct from the adrenal counterpart.

UI MeSH Term Description Entries
D008297 Male Males
D008861 Microsomes Artifactual vesicles formed from the endoplasmic reticulum when cells are disrupted. They are isolated by differential centrifugation and are composed of three structural features: rough vesicles, smooth vesicles, and ribosomes. Numerous enzyme activities are associated with the microsomal fraction. (Glick, Glossary of Biochemistry and Molecular Biology, 1990; from Rieger et al., Glossary of Genetics: Classical and Molecular, 5th ed) Microsome
D008862 Microsomes, Liver Closed vesicles of fragmented endoplasmic reticulum created when liver cells or tissue are disrupted by homogenization. They may be smooth or rough. Liver Microsomes,Liver Microsome,Microsome, Liver
D008970 Molecular Weight The sum of the weight of all the atoms in a molecule. Molecular Weights,Weight, Molecular,Weights, Molecular
D010671 Phenylurea Compounds Compounds that include the amino-N-phenylamide structure. Phenylcarbamides,Phenylurea Derivatives,Compounds, Phenylurea,Derivatives, Phenylurea
D011487 Protein Conformation The characteristic 3-dimensional shape of a protein, including the secondary, supersecondary (motifs), tertiary (domains) and quaternary structure of the peptide chain. PROTEIN STRUCTURE, QUATERNARY describes the conformation assumed by multimeric proteins (aggregates of more than one polypeptide chain). Conformation, Protein,Conformations, Protein,Protein Conformations
D002785 Sterol O-Acyltransferase An enzyme that catalyzes the formation of cholesterol esters by the direct transfer of the fatty acid group from a fatty acyl CoA derivative. This enzyme has been found in the adrenal gland, gonads, liver, intestinal mucosa, and aorta of many mammalian species. EC 2.3.1.26. Acyl-CoA-Cholesterol Acyltransferase,Cholesterol Acyltransferase,Cholesterol Esterifying Enzyme,Acyl CoA Cholesterol Acyltransferase,Acyltransferase, Acyl-CoA-Cholesterol,Acyltransferase, Cholesterol,Enzyme, Cholesterol Esterifying,Esterifying Enzyme, Cholesterol,O-Acyltransferase, Sterol,Sterol O Acyltransferase
D003432 Cross-Linking Reagents Reagents with two reactive groups, usually at opposite ends of the molecule, that are capable of reacting with and thereby forming bridges between side chains of amino acids in proteins; the locations of naturally reactive areas within proteins can thereby be identified; may also be used for other macromolecules, like glycoproteins, nucleic acids, or other. Bifunctional Reagent,Bifunctional Reagents,Cross Linking Reagent,Crosslinking Reagent,Cross Linking Reagents,Crosslinking Reagents,Linking Reagent, Cross,Linking Reagents, Cross,Reagent, Bifunctional,Reagent, Cross Linking,Reagent, Crosslinking,Reagents, Bifunctional,Reagents, Cross Linking,Reagents, Cross-Linking,Reagents, Crosslinking
D003510 Cyclohexanes Six-carbon alicyclic hydrocarbons.
D004791 Enzyme Inhibitors Compounds or agents that combine with an enzyme in such a manner as to prevent the normal substrate-enzyme combination and the catalytic reaction. Enzyme Inhibitor,Inhibitor, Enzyme,Inhibitors, Enzyme

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