一个单一的二次球体氨基酸替代是如何导致减少中点潜在变化数百毫伏的?
Emine Yikilmaz1, Jason Porta, Laurie E Grove
1Department of Chemistry, University of Kentucky, Lexington, KY 40506-0055, USA.
Journal of the American Chemical Society
|July 14, 2007
概括
超氧化脱酶 (SOD) 酶的活性是由其金属离子调节的.
科学领域:
- 生物化学 生物化学
- 酶学 是一种酶学.
- 蛋白质工程是指蛋白质工程.
背景情况:
- 超氧化脱酶 (SOD) 酶催化超氧化基的不成比例.
- 活性位点金属离子的降低中点潜力 (Em) 对SOD催化活性至关重要.
- 大肠杆菌含铁的SOD (FeSOD) 使用Fe3+/Fe2+氧化还原对,受质子事件的影响.
研究的目的:
- 调查大肠杆菌FeSOD突变体中Em升高的结构和电子基础.
- 了解保存的Gln残留中的突变如何影响FeSOD的氧化还原特性.
- 阐明了氧化还原合质子转移在调节酶活性中的作用.
主要方法:
- 位点导向的突变发生 (Gln到His,Gln到Glu).
- 光谱技术:EPR,MCD和NMR. 这些技术包括:
- 射线晶体学 (1.1 Å 分辨率).
- 密度函数理论 (DFT) 的计算.
主要成果:
- 突变Q69H和Q69E显著提高了FeSOD的Em.
- 谱学数据显示,这两种突变物具有类似于WT的活性部位电子结构.
- Q69E-FeSOD在减少状态下显示了变化的质子转移动态和键.
- Q69E-Fe2+SOD的晶体结构显示了Glu69.9对H键的强烈接受.
结论:
- 蛋白质矩阵通过氧化还原合质子转移显著影响FeSOD的Em.
- 在Q69E-FeSOD中改变的键和质子转移解释了Em的升高.
- 这些发现支持了金属酶中蛋白质介导的氧化还原调节假设.
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