基酶活性位点的可逆质子静止状态
Christine N Morrison1, Thomas Spatzal1, Douglas C Rees1
1Division of Chemistry and Chemical Engineering and ‡Howard Hughes Medical Institute, California Institute of Technology , Pasadena, California 91125, United States.
Journal of the American Chemical Society
|July 11, 2017
概括
酶铁 (MoFe) 蛋白在低pH下发生可逆的结构和电子变化,表明活性位点的质子化. 这些发现揭示了固的机制.
科学领域:
- 生物化学
- 结构生物学
- 生物有机化学
背景情况:
- 酶活性位点的质子化对于基质减少至关重要.
- 了解这些质子状态是阐明固定机制的关键.
研究的目的:
- 描述酶铁 (MoFe) 蛋白的低pH行为.
- 研究与质子化相关的结构和电子变化.
主要方法:
- 使用X射线结晶学和电子磁共振 (EPR) 光谱学.
- 在pH值4. 5 - 8下对来自Azotobacter vinelandii (Av) 和Clostridium pasteurianum (Cp) 的MoFe蛋白进行了研究.
主要成果:
- 低pH (4.5-6) 诱导侧链在FeMo辅因子周围重新定位.
- EPR光谱显示了一个新的S = 3/2旋转系统,类似于催化循环状态.
- 确定了FeMo辅因子带硫 (S3A或S5A) 的潜在质子位点.
- 观察到的结构和电子变化是相关的和可逆的.
- 在Av和CpMoFe蛋白之间发现了结构重组的差异.
结论:
- 可能发生静止酶状态的可逆质子化 (称为E0H+).
- 结构和电子变化为固提供了机械洞察力.
- 研究多种酶物种突出显示了活点质子化机制的变化.
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