Chemoproteomic Profiling Reveals the Mechanism of Bile Acid Tolerance in Bacteria
Chemoproteomic Profiling Reveals the Mechanism of Bile Acid Tolerance in Bacteria
复制标题
化学蛋白质组学分析揭示细菌胆汁酸耐受机制
DOI：
10.1021/acschembio.2c00286
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发表时间：
2022-09-01
影响因子：
4
通讯作者：
Wang，Chu
中科院分区：
文献类型：
--
作者：
Liu，Biwei;Zhuang，Shentian;Wang，Chu
胆汁酸是一类具有重要功能的内源性代谢产物。作为两亲性分子，BA具有很强的抗菌作用，防止肠道微生物群的过度生长并防御病原体的入侵。然而，一些致病病原体可以生存的BA压力和知识是有限的，他们如何发展BA的耐受性。在这项工作中，我们应用定量化学蛋白质组学策略来分析细菌中BA相互作用蛋白，旨在发现BA的传感途径。使用可点击和光亲和BA探针与定量质谱，我们确定了一系列的组氨酸激酶（HK）的双组分系统（TCS）在细菌中作为新的结合目标BA。基因筛选显示，敲除一种特异性HK，EnvZ，使细菌对BA具有显著的敏感性。进一步的生物化学和遗传学实验表明，BA与EnvZ中的特定口袋结合，并激活下游信号通路，以帮助BA从细菌中流出，从而导致BA耐受。总的来说，我们的数据表明，EnvZ是细菌中BA的一种新型传感器，其相关的TCS信号通路在介导细菌BA耐受性中起着关键作用，这为对抗BA耐受性病原体提供了新的机会。
Bile acids (BAs) are a class of endogenous metabolites with important functions. As amphipathic molecules, BAs have strong antibacterial effects, preventing overgrowth of the gut microbiota and defending the invasion of pathogens. However, some disease-causing pathogens can survive the BA stress and knowledge is limited about how they develop BA tolerance. In this work, we applied a quantitative chemoproteomic strategy to profile BA-interacting proteins in bacteria, aiming to discover the sensing pathway of BAs. Using a clickable and photo-affinity BA probe with quantitative mass spectrometry, we identified a list of histidine kinases (HKs) of the two-component systems (TCS) in bacteria as the novel binding targets of BA. Genetic screening revealed that knocking out one specific HK, EnvZ, renders bacteria with significant sensitivity to BA. Further biochemical and genetic experiments demonstrated that BA binds to a specific pocket in EnvZ and activates a downstream signaling pathway to help efflux of BA from bacteria, resulting in BA tolerance. Collectively, our data revealed that EnvZ is a novel sensor of BA in bacteria and its associated TCS signaling pathway plays a critical role in mediating bacterial BA tolerance, which opens new opportunities to combat BA-tolerating pathogens.