title: Purification, characterization and modulation of a microsomal carboxylesterase in rat liver for the hydrolysis of acyl-CoA.
abstract: A carboxylesterase containing long-chain acyl-CoA hydrolase activity was purified to apparent homogeneity from rat liver microsomes. Palmitoyl-CoA was the most preferred substrate, followed by stearoyl-CoA and oleoyl-CoA. Arachidonoyl-CoA, linoleoyl-CoA and acetyl-CoA were not hydrolysed by the enzyme. The purified enzyme had no activity on the hydrolysis of phospholipids and neutral lipids. The molecular mass of the enzyme was found to be 56 kDa by SDS/PAGE and 64 kDa by gel-filtration chromatography. On isoelectric focusing, the purified enzyme behaved like the ES-4 type, with a pI of 6.15. Determination of the amino acid sequence revealed that its N-terminal sequence is 100% homologous with the only other known N-terminal sequence for a rat carboxylesterase isoenzyme (ES-10). Enzyme activity was inhibited by lysophosphatidic acid and activated by lysophosphatidylcholine. The modulation of enzyme activity by these lysophospholipids might represent a plausible mechanism for the physiological control of acyl-CoA concentrations.
authors.affiliation: Department of Biochemistry & Molecular Biology, University of Manitoba, Winnipeg, Canada.
authors.forename: J J
authors.initials: JJ
authors.lastname: Mukherjee
authors.forename: F T
authors.initials: FT
authors.lastname: Jay
authors.forename: P C
authors.initials: PC
authors.lastname: Choy
mesh_terms: D000214:Acyl Coenzyme A / Q000378:metabolism*; D000818:Animals; D043182:Carboxylesterase; D002265:Carboxylic Ester Hydrolases / Q000187:drug effects / Q000378:metabolism*; D004789:Enzyme Activation; D006868:Hydrolysis; D007526:Isoelectric Point; D007700:Kinetics; D008862:Microsomes, Liver / Q000201:enzymology*; D008970:Molecular Weight; D010171:Palmitoyl Coenzyme A / Q000378:metabolism; D010170:Palmitoyl-CoA Hydrolase / Q000187:drug effects / Q000378:metabolism; D010743:Phospholipids / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D013379:Substrate Specificity
chemical_list: D000214:Acyl Coenzyme A; D010743:Phospholipids; D010171:Palmitoyl Coenzyme A; D002265:Carboxylic Ester Hydrolases; D043182:Carboxylesterase; D010170:Palmitoyl-CoA Hydrolase