共价领导探测器揭示了 ribozomal 生物合成中的外集群酶
Lan Wang1, Boning Wang1, Ying Wang1
1State Key Laboratory of Coordination Chemistry, Chemistry and Biomedicine Innovation Center of Nanjing University, Jiangsu Key Laboratory of Advanced Organic Materials, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210023, China.
Journal of the American Chemical Society
|March 16, 2026
概括
这项研究引入了一种新的化学-蛋白质学方法,使用领先 (LP) 探针来发现参与核糖体合成和转化后修饰 (RiPP) 途径的酶. 该技术成功地识别了已知基因集群之外的酶,揭示了新的跨集群活动.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 合成生物学 合成生物学
背景情况:
- 基于基因组的分析已经推进了对核糖体合成和翻译后修饰 (RiPPs) 途径的发现.
- 然而,这些方法往往错过了编码在正规生物合成基因集群之外的酶.
- 领袖 (LPs) 对于RiPP酶基质识别至关重要.
研究的目的:
- 从本地蛋白质组中开发一种化学-蛋白质组策略,直接识别LP结合酶.
- 为了利用领导 (LPs) 来实现 RiPP 酶的基质导向发现.
- 为了研究除了正规生物合成基因集群之外的酶参与.
主要方法:
- 开发了一个基质导向的化学-蛋白质战略,使用共价LP探头.
- 使用带有光交叉链接器的LP探针,在LP识别部位激活酶.
- 在模型系统 (LctA-LctM,PatE-LynD) 和*Streptomyces sparsogenes*上应用了该策略.
主要成果:
- 证明LP探头保持了酶催化能力.
- 成功识别了在*Streptomyces sparsogenes*中的正规生物合成基因集群之外编码的兰氏合成酶.
- 在多个RiPP途径中揭示了分级的交叉集群酶活性.
结论:
- 建立了一个概念验证平台,用于LP结合RiPP酶的蛋白质水平发现.
- 突出了跨集群的酶交叉作为RiPPs结构多样化的机制.
- 提供了一种新的方法来揭示RiPP生物合成的完整酶机制.
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