在2-Oxoglutarate依赖氧酶,TauD中的大型氧联重组的结构起源
Christopher W John1, Robert P Hausinger2,3, Denis A Proshlyakov1
1Department of Chemistry , Michigan State University , East Lansing , Michigan 48824 , United States.
Journal of the American Chemical Society
|September 3, 2019
概括
与氨酸酸酸酶 (TauD) 等2氧格酸盐 (2OG) 依赖的氧化酶中氧化潜力的调整与酶结构变化有关. 调查活跃部位的变体揭示了影响这种动态过程的关键残留物.
科学领域:
- 生物化学和分子生物学
- 酵素学
- 生物有机化学
背景情况:
- 2-Oxoglutarate (2OG) 依赖氧酶是催化C-H键激活的关键酶.
- 对于这些酶来说,了解瞬态中间体的机制和热力学特性至关重要.
- 之前对氨酸酶 (TauD) 的研究表明其氧化还原潜力与酶构成之间存在联系.
研究的目的:
- 调查TauD中氧化还原连接重组的结构起源.
- 确定金属结合残留物对TauD氧化还原潜力的动态调节的贡献.
- 在其他2OG-依赖氧酶中探索这些氧化还原连接的构造变化的普遍性.
主要方法:
- 光谱电化学,包括时间依赖的氧化还原定位,瞬态光学吸收和红外光谱电化学.
- 局部定向的突变产生TauD的活性局部变体 (H99A,D101Q,H255Q).
- 野生型TauD及其变体之间的氧化还原特性和构造变化的比较.
主要成果:
- 野生型的TauD表现出与形状变化相关的显著的氧化还原电位变化 (468 mV).
- 活性位点变异表明金属配体替代改变了这种重组的动力学和热力学.
- H99A变异显示出最大的氧化还原变化,表明H99的氧化还原合质子对于高氧化还原潜力至关重要.
结论:
- 与氧化物相关的形状变化是TauD的功能的一部分,涉及H99,D101和H255等特定残留物.
- 动态结构灵活性和热力学调节可能是2OG依赖氧酶家族的共同特征.
- 这项研究提供了这些重要的酶激活C-H键的机制基础.
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