在血依赖型氨酸酶中C-H和C-F键激活之间的分区的分子推理
Yifan Wang1, Ian Davis1, Inchul Shin1
1Department of Chemistry, The University of Texas at San Antonio, One UTSA Circle, Texas 78249, United States.
Journal of the American Chemical Society
|March 18, 2021
概括
血依赖的l- 铁酶 (TyrHs) 显示双重的C-H和C-F键激活. 基质导向决定了该新酶家族中的产品分布.
科学领域:
- 生物化学
- 酵素学
- 结构生物学
背景情况:
- 依赖血红素的l- 铁酶 (TyrHs) 是一个与非血红素铁酶不同的新型酶家族.
- TyrHs具有双重反应性,可以分离C-H和C-F键,特别是与3--l-tyrosine (3-F-Tyr).
- 这种双重反应背后的机制在很大程度上仍未被探索.
研究的目的:
- 从功能和结构上描述一个来自*Streptomyces sclerotialus* (SsTyrH) 的新 TyrH.
- 通过依赖血红素的TyrH来阐明双C-H和C-F键激活的分子基础.
- 确定影响TyrH催化反应产品分布的因素.
主要方法:
- 在高分辨率 (1.89 Å,1.68 Å,1.58 Å) 确定酶基质和中间结构的X射线晶体.
- 定位基因突变,以研究活性基因残留物的作用,例如His88.
- 谱分析以表征反应中间体,包括铁氧中间体 (化合物0).
主要成果:
- 与l- 铁复合的SsTyrH晶体结构揭示了电子转移启动C-H键化.
- 突变检测证实了活性部位的催化作用 (His88).
- 使用3F轮胎的结构显示基板取向是产品分布的主要决定因素 (7:3比).
- 在3-F-Tyr催化过程中观察到和结构性表征了一种铁-氧中间体 (化合物0).
结论:
- 这项研究提供了对血红素依赖的TyrH功能的第一个分子见解.
- 基质结合方向是控制C-H和C-F结合激活之间的关键因素.
- 对这种新型酶家族的结构和机制的理解已取得显著的进展.
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