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培养的中国仓鼠卵巢细胞中过氧化物酶体的特性分析。

Characterization of peroxisomes in Chinese hamster ovary cells in culture.

作者信息

Santos M J, Garrido J, Oliver C, Robbins A R, Leighton F

出版信息

Exp Cell Res. 1985 Nov;161(1):189-98. doi: 10.1016/0014-4827(85)90503-8.

Abstract

In order to explore the potential value of Chinese hamster ovary (CHO) cells for the isolation of peroxisomal mutants defective in the peroxisomal fatty acid oxidation system, some characteristics of their peroxisomes were studied. Catalase was detected biochemically and histochemically in peroxisome-like particles in cells or in subcellular fractions prepared by differential centrifugation or isopyknic equilibrium in Percoll or Metrizamide with catalase in the high density fractions of the isopyknic equilibrium gradients. By oxidation system, exhibited an unusually high specific activity, 2.46 +/- 1.09 mU/mg protein, in CHO cell homogenates, a value comparable to that of rat liver. This enzyme copurifies with catalase in the high density fractions of the isopycnic equilibrium gradients. By analogy with other cell types and from the ultrastructural analysis, it is concluded that these enzymes are contained in peroxisomes. These findings support the value of CHO cells for studies of peroxisomal function and organization.

摘要

为了探索中国仓鼠卵巢(CHO)细胞在分离过氧化物酶体脂肪酸氧化系统缺陷的过氧化物酶体突变体方面的潜在价值,对其过氧化物酶体的一些特性进行了研究。通过生化和组织化学方法在细胞中的过氧化物酶体样颗粒中或通过差速离心或在Percoll或Metrizamide中进行等密度平衡制备的亚细胞组分中检测到过氧化氢酶,在等密度平衡梯度的高密度组分中过氧化氢酶活性较高。通过氧化系统,在CHO细胞匀浆中表现出异常高的比活性,为2.46±1.09 mU/mg蛋白质,该值与大鼠肝脏相当。该酶在等密度平衡梯度的高密度组分中与过氧化氢酶共纯化。通过与其他细胞类型类比并根据超微结构分析,得出这些酶存在于过氧化物酶体中的结论。这些发现支持了CHO细胞在过氧化物酶体功能和组织研究中的价值。

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