希斯蒂丁定向调节新金属酶活性位点的结构和动态
Matthew R Ross1, Aaron M White1, Fangting Yu1
1Department of Chemistry, University of Michigan, Ann Arbor, Michigan 48109, United States.
Journal of the American Chemical Society
|August 7, 2015
概括
这项研究使用二维红外光谱来观察新型金属酶的超快结构变化. 的振动激发显示出快速的重组和缓慢的放松,受胺侧链和蛋白质静电的影响.
科学领域:
- 生物物理化学
- 酶催化
- 光谱学
背景情况:
- 新型金属酶为研究酶机制提供可调节的活性位点.
- 与铜 (Cu) 结合的一氧化碳 (CO) 作为金属酶活性部位的振动探针.
- 了解蛋白质静电在酶功能中的作用对于生物工程至关重要.
研究的目的:
- 使用二维红外光谱来研究新金属酶活性部位的超快动态.
- 阐明CO连接体振动和蛋白质结构重组之间的合机制.
- 探索蛋白质静电如何影响金属酶活性部位的动态和功能.
主要方法:
- 两维红外 (2D IR) 光谱仪用于监测超快的动态.
- 量子力学 (QM) 和QM/MM计算以确定合坐标.
- 对金属酶活性部位的结构和静电特性进行分析.
主要成果:
- 观察到CO振动激发的超快 (2 ps) 失衡结构重组.
- 识别了金属-CO振动特征的较慢的振动放松 (∼40 ps).
- 计算显示二面角变化控制CO延伸曲折合,由蛋白质静电学调节.
结论:
- 由histidine侧链转导的蛋白质静电,显著影响金属酶活性部位的潜在能量表面.
- 长距离静电相互作用可以设计为微调酶功能.
- 这项工作为功能性新型金属酶的设计原则提供了洞察力.
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