사람 양막(Aminion)에서 Oxygen Radical과 관계된 효소의 분포 및 활성도에 관한 연구

Abstract

Distribution and activity of superoxide dismutase, catalase, peroxidase and xanthine oxidase were investigated in human amnion. Human amnion was fractionated by differential centrifugation into mitochondrial and cytosolic fraction. Cytosolic superoxide dismutase was purified by ammonium sulfate precipitation, treatment with ethanol-chloroform mixture, and DEAE-cellulose column chromatography. The results were summarized as 1911ows; 1. The activity of superoxide dismutase in homogenate was found to be 20.4 units/g of wet tissue. The distribution of superoxide dismutase in the subcellular organelle showed that 94.6% of the activity was in cytosolic fraction and 5.4% in mitochondrial fraction. 2. The cytosolic superoxide dismutase was purified approximately 22 fo1d. Cyanide at 0.5mM and 5.OmM inhibited the activity of cytosolic superoxide dismutase 15.2% and 82.1%, respectively. But the activity of mitochondrial superoxide dlsmutase was inhibited about 24.4% by 1mM and 5mM cyanide. 3. The activity of catalase in homogenate was found to be 607.5 units/ml enzyme and the relative activity of cytosolic and mitochondrial catalase was 86.4% and 13.6%, respectively. 4. The activity of peroxidase in homogenate was found to be 157.4 units/ml enzyme and the relative activity of cytosolic and mitochondrial peroxidase was 83.8% and 16.2%, respectively. 5. The activity of xanthine oxidase in homogenate was found to be 3.5 units/ml enzyme and the relative activity of cytosolic and mitochondrial xanthine oridase was 66.1% and 33.9%, respectively.