Yoshioka H, Miyata T, Omura T
J Biochem. 1984 Apr;95(4):937-47. doi: 10.1093/oxfordjournals.jbchem.a134721.
When 1,1-di(p-chlorophenyl)-2,2-dichloroethylene (DDE) (100 mg/kg body weight) was injected into rats, the benzphetamine N-demethylation and 7-ethoxycoumarin O-deethylation activities of liver microsomes increased by 7-fold and 3-fold, respectively, at 7 days after the injection, whereas the benzo(a)pyrene 3-hydroxylation activity did not increase. The content of cytochrome P-450 in the microsomes increased 2.5-fold at 7 days. By the use of the antibodies to a phenobarbital (PB)-inducible form (P-450(PB-1)) and a 3-methylcholanthrene (MC)-inducible form (P-450(MC-1)) of cytochrome P-450, the contents of P-450(PB-1) and P-450(MC-1) in the liver microsomes of DDE-treated rats were measured. The form of cytochrome P-450 immunoprecipitable with anti-P-450(PB-1) antibodies increased by 10-fold at 7 days. A major component of cytochrome P-450 in the liver microsomes of DDE-treated rats, which was tentatively designated P-450(DDE), was purified. P-450 (DDE) was compared with P-450(PB-1), and they were found to be indistinguishable by the following criteria: 1) chromatographic behavior on aminooctyl-Sepharose 4B, hydroxyapatite, and DEAE-cellulose columns, 2) minimum molecular weight determined by SDS-polyacrylamide gel electrophoresis, 3) spectral properties, 4) immunoreactivity, 5) amino acid composition, 6) peptide mapping, 7) NH2-terminal amino acid sequence, 8) catalytic activities. We concluded that DDE and PB induce an identical form of P-450 in rat liver microsomes, although DDE is apparently very different from PB in chemical structure.
将1,1 - 二（对氯苯基）-2,2 - 二氯乙烯（DDE）（100毫克/千克体重）注射到大鼠体内后，在注射后7天，肝微粒体的苄非他明N - 去甲基化和7 - 乙氧基香豆素O - 去乙基化活性分别增加了7倍和3倍，而苯并（a）芘3 - 羟基化活性并未增加。微粒体中细胞色素P - 450的含量在7天时增加了2.5倍。利用针对细胞色素P - 450的苯巴比妥（PB）诱导型（P - 450（PB - 1））和3 - 甲基胆蒽（MC）诱导型（P - 450（MC - 1））的抗体，测定了DDE处理大鼠肝微粒体中P - 450（PB - 1）和P - 450（MC - 1）的含量。能用抗P - 450（PB - 1）抗体免疫沉淀的细胞色素P - 450形式在7天时增加了10倍。对DDE处理大鼠肝微粒体中细胞色素P - 450的一种主要成分进行了纯化，该成分暂定为P - 450（DDE）。将P - 450（DDE）与P - 450（PB - 1）进行比较，发现它们在以下标准方面无法区分：1）在氨辛基 - 琼脂糖4B、羟基磷灰石和DEAE - 纤维素柱上的色谱行为；2）通过SDS - 聚丙烯酰胺凝胶电泳测定的最小分子量；3）光谱特性；4）免疫反应性；5）氨基酸组成；6）肽图谱；7）NH2 - 末端氨基酸序列；8）催化活性。我们得出结论，尽管DDE在化学结构上明显与PB非常不同，但DDE和PB在大鼠肝微粒体中诱导出相同形式的P - 450。
