在完整的,由ATR-FTIR光谱学研究的质子转位型原酶中,蛋白质和尼古丁胺二核酸之间的分子识别
Masayo Iwaki1, Nick P J Cotton, Philip G Quirk
1School of Biosciences, University of Birmingham, UK.
Journal of the American Chemical Society
|February 24, 2006
概括
减弱总反射率-里埃转换红外光谱 (ATR-FTIR) 揭示了尼古丁胺二核酸如何与转化酶结合. 结合改变了蛋白质结构和二核酸环境,观察到NADP (H) 与NADP (H) 的明显变化.
科学领域:
- 生物化学 生物化学
- 频谱学是一种光谱学.
- 结构生物学 结构生物学
背景情况:
- 晶酶酶对于细胞的氧化还原平衡至关重要.
- 了解尼古丁胺二核酸结合是阐明酶机制的关键.
研究的目的:
- 为了研究尼古丁胺二核酸 (NADP+,NADPH,NAD+,NADH) 与大肠杆菌转酶的结合.
- 使用ATR-FTIR光谱学来描述核酸结合后的结构和环境变化.
主要方法:
- 使用了减弱总反射率-里埃变换红外光谱法 (ATR-FTIR).
- 没有洗剂的过酸酶被固定成薄膜.
- 光谱记录在H2O和D2O中,有和没有各种二核酸.
主要成果:
- 红外光谱变化表明二核酸的直接结合以及蛋白质结构的改变.
- 对NADPH的解离常数估计约为5μM.
- 结合诱导蛋白质二次结构 (胺基I/II带) 对NADP (H) 与NADP (H) 的明显变化.
- 结合核酸的特定红外波段显示出移位和化,表明运动受限和改变的微环境.
结论:
- 通过ATR-FTIR光谱,可以了解跨酶-二核酸相互作用.
- NADP (H) 结合似乎扰乱了蛋白质骨干的埋藏区域,并产生碳酸残留物.
- 与NADP (H) 相比,NADP (H) 存在不同的结合模式,影响蛋白质结构的不同.
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