再组合表达的hUGDH的冷-EM结构揭示了一个隐藏的,替代的菌抑制剂
John H O'Brien1, Renuka Kadirvelraj1, Po-Sen Tseng2
1Department of Biochemistry & Molecular Biology, University of Georgia, Athens, Georgia 30602, United States.
Biochemistry
|December 16, 2024
概括
人类UDP-葡萄糖脱酶 (hUGDH) 存在于活性和非活性状态. 我们发现UDP-4-基 (UX4O) 稳定了非活性状态,解决了hUGDH全机制研究中的差异.
科学领域:
- 生物化学 生化学
- 酶学 是一种酶学.
- 结构生物学 结构生物学
背景情况:
- 人类UDP-葡萄糖脱酶 (hUGDH) 对于药物代谢至关重要.
- hUGDH存在于活性 (E) 和非活性 (EΩ) 状态之间的平衡状态,受到UDP-Xylose (UDP-Xyl) 等全抑制剂的影响.
- 以前的研究提出了关于未结合的hUGDH的首选状态的相互矛盾的证据.
研究的目的:
- 为了解决hUGDH的全性机制中的差异.
- 为了研究UDP-4-keto-xylose (UX4O) 在hUGDH构成状态中的作用.
- 为了确定UX4O作为抑制剂的生理相关性.
主要方法:
- 低温电子显微镜 (cryo-EM) 用于可视化hUGDH结构.
- 生物化学试验分析酶动力学和抑制.
- 从大肠杆菌 (Escherichia coli) 中净化复合的hUGDH.
主要成果:
- 重组hUGDH与UX4O共净化,稳定了非活性EΩ状态.
- 移除UX4O将hUGDH组合转移到活动E状态.
- 进度曲线分析证实,在UX4O去除后,无结合的hUGDH没有滞后.
- hUGDH对UDP-Xyl和UX4O具有类似的结合亲缘关系.
结论:
- UX4O被确定为影响hUGDH构造状态的关键因素.
- 这些发现支持未结合的hUGDH向活性状态的转变.
- 建议UX4O作为细菌UDP-葡萄糖脱酶的潜在生理抑制剂.
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