Immunotitration of 3-hydroxy-3-methylglutaryl-coenzyme A reductase in various physiological states.
Immunotitration of 3-hydroxy-3-methylglutaryl-coenzyme A reductase in various physiological states.
复制标题
各种生理状态下 3-羟基-3-甲基戊二酰辅酶 A 还原酶的免疫滴定。
DOI：
10.1073/pnas.76.8.3834
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发表时间：
1979
影响因子：
11.1
通讯作者：
T. Scallen
中科院分区：
文献类型：
--
作者：
J. E. Hardgrave;R. Heller;M. Herrera;T. Scallen
本文报道了胆固醇生物合成的主要调节酶3-羟基-3-甲基戊二酰辅酶A（HMG-CoA）还原酶[甲羟戊酸：NADP+氧化还原酶[CoA-酰化]，EC 1.1.1.34]的免疫滴定法。该技术提供的优点是，通过使用从粗肝微粒体到纯化酶的酶样品，可以快速测量不同生理状态下酶浓度和酶活性的相对变化。关于HMG-CoA还原酶的昼夜节律，在接近黑暗期中期（D 4.5）处死的动物与接近光照期中期（L 4.5）处死的动物之间观察到的主要差异在于存在的HMG-CoA还原酶浓度，而不是酶活性，在接近黑暗期中期发现了更多的酶。考来烯胺处理导致HMG-CoA还原酶的浓度增加和催化活性更高的酶。另一方面，胆固醇喂养导致HMG-CoA还原酶浓度降低和催化活性降低。
The immunotitration of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase [mevalonate:NADP+ oxidoreductase [CoA-acylating), EC 1.1.1.34], the major regulatory enzyme in cholesterol biosynthesis, by HMG-CoA reductase antiserum is described. This technique provides the advantage that one can rapidly measure relative changes in both enzyme concentration and enzyme activity in different physiological states by using enzyme samples all the way from crude liver microsomes to purified enzyme. Regarding the diurnal rhythm of HMG-CoA reductase, the major difference noted between animals killed near the middle of the dark period (D 4.5) compared to animals killed near the middle of the light period (L 4.5) was in the concentrations of HMG-CoA reductase present, rather than in the activity of the enzyme, with substantially more enzyme found near mid-dark. Cholestyramine treatment resulted in both an increased concentration of HMG-CoA reductase and a catalytically more active enzyme. On the other hand, cholesterol feeding resulted in both a decreased concentration of HMG-CoA reductase and a catalytically less active enzyme.