一种用于过氧化物酶体酰基辅酶A氧化酶的灵敏分光光度测定法。
A sensitive spectrophotometric assay for peroxisomal acyl-CoA oxidase.
作者信息
Small G M, Burdett K, Connock M J
出版信息
Biochem J. 1985 Apr 1;227(1):205-10. doi: 10.1042/bj2270205.
Abstract
A simple spectrophotometric assay was developed for peroxisomal fatty acyl-CoA oxidase activity. The assay, based on the H2O2-dependent oxidation of leuco-dichlorofluorescein catalysed by exogenous peroxidase, is more sensitive than methods previously described. By using mouse liver samples, cofactor requirements were assessed and a linear relationship was demonstrated between dye oxidation and enzyme concentration. By using this assay on subcellular fractions, palmitoyl-CoA oxidase activity was localized for the first time in microperoxisomes of rat intestine. The assay was also adapted to measure D-amino acid oxidase activity, demonstrating the versatility of this method for measuring activity of other H2O2-producing oxidases.
摘要
开发了一种用于检测过氧化物酶体脂肪酰辅酶A氧化酶活性的简单分光光度法。该检测方法基于外源过氧化物酶催化的依赖于H2O2的无色二氯荧光素氧化反应，比先前描述的方法更灵敏。通过使用小鼠肝脏样本，评估了辅因子需求，并证明了染料氧化与酶浓度之间的线性关系。通过对亚细胞组分使用该检测方法，首次在大鼠肠道的微小过氧化物酶体中定位了棕榈酰辅酶A氧化酶活性。该检测方法还适用于测量D-氨基酸氧化酶活性，证明了该方法在测量其他产生H2O2的氧化酶活性方面的通用性。
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