Electron paramagnetic resonance (EPR) studies on hydrogenase-1 (HYD1) purified from a mutant strain (AP6) of Escherichia coli enhanced in HYD1.
Electron paramagnetic resonance (EPR) studies on hydrogenase-1 (HYD1) purified from a mutant strain (AP6) of Escherichia coli enhanced in HYD1.
复制标题
对从 HYD1 增强的大肠杆菌突变株 (AP6) 中纯化的氢化酶 1 (HYD1) 进行电子顺磁共振 (EPR) 研究。
DOI：
10.1006/bbrc.1996.1491
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发表时间：
1996
影响因子：
3.1
通讯作者：
DerVartanian,DV
中科院分区：
文献类型：
--
作者：
DerVartanian,ME;Menon,NK;Przybyla,AE;PeckJr,HD;DerVartanian,DV
氢酶-1（HYD 1）过表达两倍，已从缺乏氢酶-2的大肠杆菌突变株（AP 6）中纯化至同质且具有高比活性。等离子体发射光谱表明，0.93原子的镍和11.4铁原子存在于HYD 1。EPR研究作为隔离HYD 1检测到一个复杂的3Fe-4S信号和Ni（III）物种。用氢气还原导致3Fe-4S簇和初始Ni（III）信号消失。同时，出现活化的氢化酶的EPR特征（小g = 2.19信号）。注意到检测到4Fe-4S簇信号。用连二亚硫酸钠还原HYD 1导致所有镍信号消失。4Fe-4S络合强度略有增加。在三种氧化还原状态下的EPR响应与其他已知的（NiFe）-氢化酶是一致的。
Hydrogenase-1 (HYD1), overexpressed by twofold, has been purified to homogeneity and to a high specific activity from a mutant strain (AP6) ofEscherichia coliwhich lacks hydrogenase-2. Plasma emission spectroscopy indicated that 0.93 atom of nickel and 11.4 iron atoms were present in HYD1. EPR studies on the as isolated HYD1 detected a complex 3Fe-4S signal and a Ni(III) species. Reduction with hydrogen gas caused disappearance of both the 3Fe–4S cluster and initial Ni(III) signals. At the same time the EPR signature (small g=2.19 signal) of the activated hydrogenase appeared. The detection of a 4Fe–4S cluster signal was noted. Reduction of HYD1 with sodium dithionite caused all nickel signals to disappear. The 4Fe–4S complex intensity was slightly increased. The EPR responses in the three oxidation-reduction states are consistent with other known (NiFe)-hydrogenases.