Partial purification and properties of long-chain acyl-CoA hydrolase from rat brain cytosol.
Long-chain acyl-CoA hydrolase (EC 3.1.2.2) has been partially purified from the 100,000 X g supernatant fraction of rat brain tissue. The purification procedure included chromatography on gel filtration media, DEAE-cellulose, CM-cellulose, and hydroxyapatite. The partially purified enzyme had a specific activity of 7.1 mumol/min-mg, and when analyzed by polyacrylamide gel electrophoresis, revealed one major and three minor bands of protein in the presence of dodecyl sulfate and two major bands of protein in the absence of dodecyl sulfate. The enzyme had a molecular weight of 65,000 and showed no evidence of aggregated or dissociated forms. The highest catalytic activity was exhibited with palmitoyl-CoA and oleoyl-CoA as substrates. Lower activity was found with decanoyl-CoA as the substrate and little or no activity was found with acetyl-CoA, malonyl-CoA, butyryl-CoA, or acetoacetyl-CoA. The enzyme was inhibited by CoA, various metal ions, including Mn2+, Mg2+ and Ca2+, and by bovine serum albumin. Heating the enzyme produced a loss of activity which corresponded to a first-order kinetic process, the rate of which was independent of the choice of substrate used to measure enzyme activity. This finding supports the idea that the purification procedure yields a single species of long-chain acyl-CoA hydrolase.
D000214:Acyl Coenzyme A / Q000378:metabolism; D000818:Animals; D001921:Brain / Q000201:enzymology*; D003600:Cytosol / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D007700:Kinetics; D008970:Molecular Weight; D010171:Palmitoyl Coenzyme A / Q000378:metabolism; D010170:Palmitoyl-CoA Hydrolase / Q000302:isolation & purification* / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D013053:Spectrophotometry; D013869:Thiolester Hydrolases / Q000302:isolation & purification*
D000214:Acyl Coenzyme A; C017585:oleoyl-coenzyme A; D010171:Palmitoyl Coenzyme A; D013869:Thiolester Hydrolases; D010170:Palmitoyl-CoA Hydrolase