Pyruvate formate-lyase-activating enzyme:: Strictly anaerobic isolation yields active enzyme containing a [3Fe-4S]+ cluster
Pyruvate formate-lyase-activating enzyme:: Strictly anaerobic isolation yields active enzyme containing a [3Fe-4S]+ cluster
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DOI：
10.1006/bbrc.2000.2313
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发表时间：
2000-03-16
影响因子：
3.1
通讯作者：
Broderick, WE
中科院分区：
文献类型：
--
作者：
Broderick, JB;Henshaw, TF;Broderick, WE
来自大肠杆菌（E. coli）的丙酮酸甲酸裂解酶激活酶（PFL- ae）催化丙酮酸甲酸裂解酶（PFL） Gly734中一个氢原子的立体特异性提取，该反应严格依赖于共底物s -腺苷-l -蛋氨酸（AdoMet）。虽然Pn-AE是一种铁依赖性酶，但由于金属中心的氧敏感性和不稳定性，分离金属中心完整的酶被证明是困难的。我们在这里报道了在非变性、严格厌氧条件下首次分离到的PFL-AE。铁和硫化物分析以及W-visible， EPR和共振拉曼数据支持纯化酶中存在[3Fe-4S]（+）簇。分离的天然酶，而不是猿酶，在没有添加铁的情况下表现出很高的比活性（31 U/mg），表明天然簇对酶活性是必要的和充分的，(C)学术出版社。
Pyruvate formate-lyase-activating enzyme (PFL-AE) from Escherichia coli (E. coli) catalyzes the stereospecific abstraction of a hydrogen atom from Gly734 of pyruvate formate-lyase (PFL) in a reaction that is strictly dependent on the cosubstrate S-adenosyl-L-methionine (AdoMet). Although Pn-AE is an iron-dependent enzyme, isolation of the enzyme with its metal center intact has proven difficult due to the oxygen sensitivity and lability of the metal center. We report here the first isolation of PFL-AE under nondenaturing, strictly anaerobic conditions. Iron and sulfide analysis as well as W-visible, EPR, and resonance Raman data support the presence of a [3Fe-4S](+) cluster in the purified enzyme. The isolated native enzyme, but not ape-enzyme, exhibits a high specific activity (31 U/mg) in the absence of added iron, indicating that the native cluster is necessary and sufficient for enzymatic activity, (C) Academic Press.