在核酸还原酶内进行形态动态基因转移
Brandon L Greene1, Alexander T Taguchi2, JoAnne Stubbe2
1Department of Chemistry and Chemical Biology, Harvard University , Cambridge, Massachusetts 02138, United States.
Journal of the American Chemical Society
|October 18, 2017
概括
核酸减少酶使用基因转移进行核酸减少. 这项研究表明, 蛋白质的灵活性使得基因的转移速度更快,
科学领域:
- 生物化学
- 酵素学
- 蛋白质动力学
背景情况:
- 核酸减少酶 (RNR) 是DNA合成的必需酶.
- 1a类RNR使用涉及囊基的基转移 (RT) 机制.
- 在α2:β2子单元接口中通过质子合电子转移 (PCET) 发生基质转移.
研究的目的:
- 研究构造力学在RNR基转移中的作用.
- 描述大肠杆菌1a类RNR的R411A突变分子转移的机制.
- 通过子单元接口确定激素转移的动力学.
主要方法:
- 位点定向的突变发生 (R411A).
- 珀子抑制以安装3-氨基氨酸 (NH2Y) 作为激素陷.
- 通过HYSCORE光谱来研究被困的激素状态.
- 用于运动研究的光化学激素生成.
主要成果:
- 破坏Y731H结合的R411A突变保留了酶活性,表明了形状的灵活性.
- 在R411A突变中,Y731动态地改造H键,允许基因转移传播.
- 在R411A突变体中,基因转移依赖于形状.
- 在ns-μs时间尺度上发生了Y731的形状变化,比催化速率更快.
结论:
- 氨酸残留物的形态灵活性对于在RNR中有效的基因转移至关重要.
- 动态H键重组促进了沿着PCET途径的基质传播.
- 酶动力学是由蛋白质动力学和基质运输之间的相互作用调节的.
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