沙漠蝗（Schistocerca gregaria）中α-蜕皮激素C-20羟基化的机制。
The mechanism of C-20 hydroxylation of alpha-ecdysone in the desert locust, Schistocerca gregaria.
作者信息
Johnson P, Rees H H
出版信息
Biochem J. 1977 Dec 15;168(3):513-20. doi: 10.1042/bj1680513.
Abstract
- The C-20 hydroxylation of alpha-ecdysone to produce beta-ecdysone was investigated in the desert locust, Schistocerca gregaria. 2. alpha-Ecdysone C-20 hydroxylase activity was located primarily in the fat-body and Malpighian tubules. The properties of the hydroxylation system from Malpighian tubules investigated further. 3. The enzyme system was mitochondrial, had a pH optimum of 6.5, an apparent Km of 12.5 micron and required O2 and NADPH. 4. The activity of the hydroxylation system showed developmental variation within the fifth instar, the maximum activity corresponding to the maximum tire of endogenous moulting hormone. The significance of these results is assessed in relation to the control of the endogenous titre of beta-ecdysone. 5. The mechanism of the hydroxylation system was investigated by using known inhibitors of hydroxylation reactions such as CO, metyrapone and cyanide. 6. The CO difference spectrum of the reduced mitochondrial preparation indicated the presence of cytochrome P-450 in the preparation. 7. It concluded that the alpha-ecdysone C-20 hydroxylase system is a cytochrome P-450-deendent mono-oxygenase.
摘要
- 对沙漠蝗（Schistocerca gregaria）中α-蜕皮激素C-20位羟基化生成β-蜕皮激素的过程进行了研究。2. α-蜕皮激素C-20羟化酶活性主要位于脂肪体和马氏管中。对来自马氏管的羟基化系统的特性进行了进一步研究。3. 该酶系统存在于线粒体中，最适pH为6.5，表观Km为12.5微摩尔，需要氧气和NADPH。4. 羟基化系统的活性在五龄期内呈现发育变化，最大活性与内源性蜕皮激素的最高水平相对应。结合β-蜕皮激素内源性滴度的控制对这些结果的意义进行了评估。5. 通过使用已知的羟基化反应抑制剂如一氧化碳、甲吡酮和氰化物，对羟基化系统的机制进行了研究。6. 还原态线粒体制剂的一氧化碳差光谱表明制剂中存在细胞色素P-450。7. 得出结论：α-蜕皮激素C-20羟化酶系统是一种依赖细胞色素P-450的单加氧酶。
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