是否Mo参与酶MoFe蛋白的四电子减少 (E4) 中间体的化物结合?
Dmitriy Lukoyanov1, Zhi-Yong Yang, Dennis R Dean
1Department of Chemistry, Northwestern University, Evanston, Illinois 60208, USA.
这项研究提供了直接的证据,表明 (Mo) 不结合-铁辅因子 (FeMo-co) 的E(4) 中间状态中的化物. 铁 (Fe) 离子,而不是Mo,在催化周转过程中参与化物结合.
科学领域:
- 生物化学和生物有机化学
- 酶催化酶的催化作用
- 固定的研究 固的研究
背景情况:
- 铁辅因子 (FeMo-co) 是酶活性的核心.
- 了解FeMo-co内部的基质相互作用对于阐明固定机制至关重要.
- 之前的研究假设在催化过程中的化物结合中的作用.
研究的目的:
- 提供直接的实验证据,证明在催化循环过程中参与基质相互作用.
- 确定在FeMo-co.的E(4) 中间状态内的化物结合中的作用.
- 为了研究受影响的FeMo-co的电子和几何性质.
主要方法:
- 在阿尔贡下进行的H+降解过程中结捕获alpha-70 (((Ile) MoFe蛋白.
- 电子偏磁共振 (EPR) 和电子核双共振 (ENDOR) 光谱学.
- 用 (95) Mo进行同位素丰富,用于精确的高精度合测量.
主要成果:
- 激活为减少N(2) 的E(4) 中间状态,含有两个与FeMo-co结合的化物.
- 恩多尔测量结果显示,在E (4) 状态下,Mo的向量合系数很小,Mo是0.04以下.
- 这表明Mo不参与化物结合;只有Fe离子参与其中.
结论:
- 不直接结合化物在有催化作用的重要中间体E.
- 在FeMo-co中的铁离子负责催化周转过程中的化物结合.
- 尽管Mo不结合化物,但Mo的合反应表明它在调整FeMo-co的性质方面起着至关重要的作用.
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