首页 | 本学科首页   官方微博 | 高级检索  
   检索      


Partial purification and properties of long-chain acyl-CoA hydrolase from rat brain cytosol
Authors:Augustine Y Lin  Grace Y Sun  Ron macQuarrie
Institution:(1) Department of Chemistry and School of Medicine, University of Missouri-Kansas City, 64110 Kansas City, Missouri;(2) Sinclair Research Farm and Biochemistry Department, University of Missouri, 65201 Columbia, Missouri;(3) Present address: Department of Chemistry, University of Southern California, 90089 Los Angeles, California
Abstract:Long-chain acyl-CoA hydrolase (EC 3.1.2.2.) has been partially purified from the 100,000 × 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 mgrmol/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.
Keywords:
本文献已被 SpringerLink 等数据库收录!
设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号