Use of trypsin and lipoamidase to study the role of lipoic acid moieties in the pyruvate and alpha-ketoglutarate dehydrogenase complexes of Escherichia coli.

Use of trypsin and lipoamidase to study the role of lipoic acid moieties in the pyruvate and alpha-ketoglutarate dehydrogenase complexes of Escherichia coli.
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使用胰蛋白酶和脂酰胺酶研究硫辛酸部分在大肠杆菌的丙酮酸和 α-酮戊二酸脱氢酶复合物中的作用。

DOI:
10.1021/bi00519a007
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发表时间:
1981
期刊:
影响因子:
2.9
通讯作者:
Reed,LJ
Reed,LJ
中科院分区:
生物学3区
文献类型:
--
作者:
Stepp,LR;Bleile,DM;McRorie,DK;Pettit,FH;Reed,LJ

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Larry R.Stepp,Dennis M.Bleile,*Donald K.McRorie,Flora H.Pettit和Lester J.Reed*摘要:研究了大肠杆菌丙酮酸和酮戊二酸脱氢复合体中~3H标记的硫辛基部分被胰酶和脂酰胺酶释放与总的酶活性损失之间的关系。胰酶从二氢硫辛基转乙酰基酶和二氢硫辛基转移琥珀酰基酶的“内部”核心释放硫酰基结构域及其共价连接的硫酰基部分,而硫胺酶只释放硫酰基部分。结果表明,胰酶释放硫基结构域和脂酰胺酶释放硫酰基部分的速度比伴随的两个络合物的总活性损失更快。胰酶释放丙酮酸脱氢酶复合体中大约一半的硫辛基结构域,而对整体活性没有显著影响。提出了一个模型来解释这些和其他关于通过硫辛基部分的活性中心耦合的观察结果。大肠杆菌的丙酮酸和-酮戊二酸脱氢酶复合体由三种不同的酶组成,它们催化以下一系列反应:
Larry R. Stepp, Dennis M. Bleile,* Donald K. McRorie, Flora H. Pettit, and Lester J. Reed* abstract: The relationships between release of 3H-labeled lipoyl moieties by trypsin and lipoamidase and accompanying loss of overall enzymatic activityof the Escherichia coli py-ruvate and-ketoglutarate dehydrogenasecomplexes were studied. Trypsin releases lipoyl domains together with their covalently attached lipoyl moieties from the “inner” core of the dihydrolipoyl transacetylase and the dihydrolipoyl transsuccinylase whereas lipoamidase releases only the lipoyl moieties. The results show that release of lipoyldomains by trypsin and release of lipoyl moieties by lipoamidaseproceeded at faster rates than the accompanying loss of overall activity of the two complexes. Trypsin released about half of the lipoyl domains in the pyruvate dehydrogenase complex without significant effect on the overall activity. A model is presented to explain these andother observations on active-site coupling via lipoyl moieties. e pyruvate and-ketoglutarate dehydrogenase complexes of Escherichia coli consist of three different enzymes that catalyze the following sequence of reactions:
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