ACYL-COENZYME-A - CHOLESTEROL O-ACYLTRANSFERASE IS NOT IDENTICAL TO LIVER MICROSOMAL CARBOXYLESTERASE

Acyl-coenzyme A (CoA):cholesterol O-acyltransferase (ACAT) is responsi ble for esterification of cholesterol in the cell. The enzyme has neve r been purified, but two cDNA sequences coding for this enzyme were re cently reported. One of the sequences was identical to human liver car boxylesterase. We have used inhibitors to elucidate the relation betwe en microsomal carboxylesterase, acyl-CoA hydrolase (AGH), and ACAT act ivities in rat liver. Low concentrations of serine esterase inhibitors strongly inhibited carboxylesterase and acyl-CoA hydrolase activities but stimulated ACAT activity. At higher concentrations, ACAT activity was also inhibited. A sulfhydryl-modifying agent was found to be a po tent inhibitor of ACAT without affecting carboxylesterase activity. Si milarly, two specific ACAT inhibitors, DL-melinamide and PD 138142-15, inhibited ACAT activity but did not affect carboxylesterase or ACH ac tivities. Our data thus exclude ACAT as a liver microsomal carboxylest erase. The complex inhibition patterns observed with serine esterase i nhibitors indicate that carboxylesterases and ACHs may interfere with ACAT activity by competing for the substrate. It is obvious that final identification of ACAT requires demonstration of an active homogenous protein.