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在线无氧小角度X射线散射的开发和氧感应转录调节器的结构特征

Gabrielle Illava¹, Richard Gillilan², Nozomi Ando³

¹Department of Chemistry and Chemical Biology, Cornell University, Ithaca, New York, USA; Center for High Energy X-ray Sciences (CHEXS), Ithaca, New York, USA.

The Journal of biological chemistry

|July 13, 2023

概括

我们开发了一种无氧小角度X射线散射 (anSAXS) 系统,用于研究对氧敏感的金属酶. 这个系统提供了氧气诱导的蛋白质变化的直接结构证据,如FNR转录因子.

科学领域:

生物化学和生物物理学
结构生物学结构生物学
在X射线散射中.

背景情况:

对氧气敏感的金属酶对于基本的生物过程至关重要.
在无氧条件下在非低温温度下对这些蛋白质进行表征是具有挑战性的.
烟酸和酸盐还原 (FNR) 转录因子的活性由氧和可变性 [4Fe-4S] 集群调节.

研究的目的:

引入和验证第一个直线无氧小角度X射线散射 (anSAXS) 系统.
为了研究大肠杆菌FNR转录因子对氧气反应的结构变化.
通过结合色谱学-anSAXS.S.展示金属蛋白-DNA相互作用的研究.

主要方法:

在线无氧小角度X射线散射 (anSAXS) 系统的开发,具有批量和染色学模式.
染色学合的anSAXS用于研究FNR转录因子的寡合体相互转换.
将尺寸排除色谱-anSAXS与UV-Vis分析结合起来,以研究FNR-DNA相互作用.

主要成果:

获得了第一个直接的结构证据,证明大肠杆菌FNR二分体的氧气诱导解离.
FNR二聚体的解离与其 [4Fe-4S] 集群的降解有关.
据证明,Dimeric FNR及其 [4Fe-4S] 集群与 nrdDG 发起区的协同站点结合.

结论:

在线开发的anSAXS系统显著推进了对氧敏感金属蛋白的研究.
这种技术在无氧条件下提供了对像FNR这样的蛋白质结构动态的关键见解.
anSAXS系统为研究复杂的金属蛋白和金属蛋白-DNA相互作用提供了一个强大的新工具.

关键词:

这是一种DNA结合蛋白质.厌氧代谢的厌氧代谢铁硫蛋白质是一种铁硫蛋白质.这是一种金属蛋白质金属蛋白.氧化聚合物的一种.小角X射线散射的小角度X射线散射.结构-功能的结构-功能.转录条例转录条例转录条例

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