Mg2+ 结合于硫基酸合成酶的表面,并影响内部活性部位的化物转移
Zhen Wang1, Paul J Sapienza, Thelma Abeysinghe
1Department of Chemistry, University of Iowa, Iowa City, Iowa 52242, USA.
Journal of the American Chemical Society
|April 25, 2013
概括
离子 (Mg2+) 结合于基酸合成酶 (TSase) 的表面,稳定其结构,并将DNA合成加速7倍. 这种远程相互作用会影响酶活性,并指导新药的设计.
科学领域:
- 生物化学 生物化学
- 酶学 是一种酶学.
- 结构生物学 结构生物学
背景情况:
- 乙基胺酸合成酶 (TSase) 对于DNA合成至关重要,也是药物的关键标.
- 离子 (Mg2+) 对TSase活性的影响已知,但潜在的机制尚不清楚.
研究的目的:
- 阐明Mg2+与大肠杆菌TSase相互作用并调节其活性的机制.
- 为Mg2+结合及其对TSase功能的影响提供结构和动力学证据.
主要方法:
- 使用X射线晶体学来确定TSase与结合Mg2+的结构.
- 进行了酶动力学和核磁共振 (NMR) 放松实验,以分析Mg2+对TSase活性和动态的影响.
主要成果:
- 2+与大肠杆菌TSase的外部结合,靠近叶酸辅因子的谷氨基部分,这是这种结合的第一个结构证据.
- 结合Mg2+稳定了封闭的三元酶复合体,降低了激活和加快了约7倍的化物转移.
- 动态同位素效应保持不变,表明Mg2+促进了蛋白质运动以接近基质,而不是改变化物转移步骤本身.
结论:
- Mg2+通过远程表面相互作用调节TSase活性,影响蛋白质动力学和酶动力学.
- 了解TSase的Mg2+调节为通过向细菌TSase相互作用来设计特定物种的抗叶酸抗生素提供了见解.
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