Further studies on the submitochondrial localization of cholesterol side chain-cleaving enzyme system in hog adrenal cortex by sonic treatment.
Further studies on the submitochondrial localization of cholesterol side chain-cleaving enzyme system in hog adrenal cortex by sonic treatment.
复制标题
声波处理猪肾上腺皮质胆固醇侧链裂解酶系统软骨下定位的进一步研究。
DOI：
10.1093/oxfordjournals.jbchem.a129414
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发表时间：
1970
影响因子：
2.7
通讯作者：
Shogo Ichii
中科院分区：
文献类型：
--
作者：
Nagasumi Yago;Shigeru Kobayashi;Shigetaka Sekiyama;Hiromi Kurokawa;Yuko Iwai;Ichiro Suzuki;Shogo Ichii
1.猪肾上腺皮质线粒体，其纯度通过电子显微镜观察和通过测定微粒体类固醇21-羟化酶[EC 1.14.1. 8]，在超声处理后通过离心进行分级。对9，000 ~ 105，000×g、105，000 ~ 150，000 × g和150，000×g超声处理的线粒体上清液中的沉淀物进行酶活性和细胞色素含量测定.以琥珀酸脱氢酶[EC www.example.com ]作为内膜标记物，Amytal不敏感的NADH细胞色素c还原酶和细胞色素bs作为外膜标记物，分别将105，000 × g沉淀物确定为内膜制剂，150，000×g沉淀物确定为外膜制剂，150，000×g上清液确定为基质制剂1.3.99.1。和苹果酸脱氢酶[EC 1.1.1. 37]和苹果酸酶[EC 1.1.1. 40]对于矩阵。3.在存在或不存在外源性NADPH黄递酶[EC 1.6. 99.1]和肾上腺素能氧还蛋白，清楚地表明这些酶的活性位于内膜.丙酮处理对线粒体酶的差异提取效果不佳.目前的结果与以前的一起似乎建立本地化的P-450和胆固醇侧链裂解酶系统的羟化酶在猪肾上腺皮质的线粒体内膜。
1. Hog adrenocortical mitochondria, of which purity was assured by electron microscopic observation and by determination of the microsomal steroid 21-hydroxylase [EC 1.14.1. 8], were fractionated by centrifugation after sonication. The precipitates obtained between 9, 000 and 105, 000×g, and between 105, 000 and 150, 000×g as well as the 150, 000×g supernatant of the sonicated mitochondria were examined with respect to their enzymatic activities and cytochrome contents.2. The 105, 000Xg precipitate was identified as the inner membrane preparation, the 150, 000×g precipitate as the outer membrane preparation, and the 150, 000×g supernatant as the matrix preparation, respectively, using succinate dehydrogenase [EC 1.3.99.1] as the marker for inner membrane, the Amytal-insensitive NADH cytochrome c reductase and cytochrome bs for outer membrane, and malate dehydrogenase [EC 1.1.1. 37] and malic enzyme [EC 1.1.1. 40] for matrix.3. Incubation for the assay of hydroxylases of cholesterol side chain-cleaving enzyme system, in the presence or absence of exogenous NADPH diaphorase [EC 1.6. 99.1] and adrenodoxin, clearly demonstrated that these enzyme activities were located in the inner membrane.4. Acetone treatment of mitochondria was not effective for the differential extraction of mitochondrial enzymes.5. Present results together with the previous ones seem to establish the localization of the P-450 and hydroxylases of the cholesterol side chain-cleaving enzyme system at the inner mitochondrial membrane of the hog adrenal cortex.