调节一个先前存在的构造平衡,调整腺酸酶活性
Jörgen Ådén1, Abhinav Verma, Alexander Schug
1Department of Chemistry, Chemical Biological Center, Umeå University, SE-901 87 Umeå, Sweden.
Journal of the American Chemical Society
|September 12, 2012
概括
腺酸酶 (AK(eco)) 的结构可塑性是其酶活性的关键. 调节其构造平衡会影响周转率和基质结合,从而提供进化灵活性.
科学领域:
- 生物化学 生物化学
- 结构生物学 结构生物学
- 酶学 是一种酶学.
背景情况:
- 酶催化依赖于反应周期的结构可塑性.
- 来自大肠杆菌 (AK(eco)) 的腺酸酶存在于开放 (无活性) 和闭合 (活性) 状态.
- 在AK(eco) 中的酶周转率与其活性构成的寿命有关.
研究的目的:
- 在没有基质的情况下,研究AK{\displaystyle AK{\mathrm {eco } } 的结构平衡.
- 了解这种平衡如何影响酶催化.
- 探索突变和奥斯莫利特对AK (生态) 构造和活动的影响.
主要方法:
- 核磁共振 (NMR) 光谱学.核磁共振 (NMR) 光谱学.
- 分子动力学 (MD) 模拟.
- 局部定向的突变发生和溶剂治疗.
主要成果:
- 移除一个键将平衡转移到开放的形状,增加k(cat).
- 添加TMAO将平衡转移到封闭的形状,减少k(cat).
- 迈克利斯常数 (K(M)) 与k(cat) 变化相关,表明基质结合亲和力发生变化.
结论:
- 预先存在的构造平衡直接影响了酶催化.
- k ((cat) 和K ((M) 是相互依赖的,并受到形状群体的影响.
- AK(eco) 具有灵活性,可以根据细胞选择性压力调整其特异性常数.
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