Hepatic acyl-CoA:cholesterol acyltransferase. Development of a standard assay and determination in patients with cholesterol gallstones.
A standard assay was developed for human liver acyl-CoA:cholesterol acyltransferase (ACAT, EC 2.3.2.26) which is more sensitive than previous methods and allows accurate activity determinations on crude microsomal fractions. ACAT activity was measured in microsomes from livers of four gallstone patients and five controls. Preincubation with exogenous cholesterol produced an increase in ACAT activity in all liver samples: gallstone samples showed a mean increase of 1.8-fold, whereas non-gallstone samples showed a mean increase of 8.2-fold. The mean ACAT activity measured in the presence of exogenous cholesterol was 52.8 +/- 22.8 (n = 4) pmol . min-1 . mg-1 for gallstone samples and 82.8 +/- 13.5 (n = 4) pmol . min-1 . mg-1 for non-gallstone samples. These results suggest that patients suffering from cholesterol gallstones have a reduced ability to esterify potentially harmful free cholesterol compared with controls. They support the proposition that cholesterol gallstone formation is related to altered hepatic cholesterol metabolism.
D000214:Acyl Coenzyme A / Q000032:analysis; D000368:Aged; D000818:Animals; D002021:Buffers; D002417:Cattle; D002769:Cholelithiasis / Q000201:enzymology*; D002784:Cholesterol / Q000032:analysis*; D003902:Detergents; D004795:Enzyme Stability; D005260:Female; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008099:Liver / Q000201:enzymology*; D008297:Male; D008875:Middle Aged; D012710:Serum Albumin, Bovine / Q000032:analysis; D002785:Sterol O-Acyltransferase / Q000032:analysis*
D000214:Acyl Coenzyme A; D002021:Buffers; D003902:Detergents; C017585:oleoyl-coenzyme A; D012710:Serum Albumin, Bovine; D002784:Cholesterol; D002785:Sterol O-Acyltransferase