A pathogen-specific isotope tracing approach reveals metabolic activities and fluxes of intracellular Salmonella.

IF 7.8 1区 生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY PLoS Biology Pub Date : 2023-08-18 eCollection Date: 2023-08-01 DOI:10.1371/journal.pbio.3002198
Karin Mitosch, Martin Beyß, Prasad Phapale, Bernhard Drotleff, Katharina Nöh, Theodore Alexandrov, Kiran R Patil, Athanasios Typas
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引用次数: 3

Abstract

Pathogenic bacteria proliferating inside mammalian host cells need to rapidly adapt to the intracellular environment. How they achieve this and scavenge essential nutrients from the host has been an open question due to the difficulties in distinguishing between bacterial and host metabolites in situ. Here, we capitalized on the inability of mammalian cells to metabolize mannitol to develop a stable isotopic labeling approach to track Salmonella enterica metabolites during intracellular proliferation in host macrophage and epithelial cells. By measuring label incorporation into Salmonella metabolites with liquid chromatography-mass spectrometry (LC-MS), and combining it with metabolic modeling, we identify relevant carbon sources used by Salmonella, uncover routes of their metabolization, and quantify relative reaction rates in central carbon metabolism. Our results underline the importance of the Entner-Doudoroff pathway (EDP) and the phosphoenolpyruvate carboxylase for intracellularly proliferating Salmonella. More broadly, our metabolic labeling strategy opens novel avenues for understanding the metabolism of pathogens inside host cells.

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病原体特异性同位素追踪方法揭示了细胞内沙门氏菌的代谢活性和通量。
在哺乳动物宿主细胞内增殖的病原菌需要快速适应细胞内环境。由于难以在原位区分细菌和宿主代谢产物,它们如何实现这一点并从宿主中清除必需营养素一直是一个悬而未决的问题。在这里,我们利用哺乳动物细胞不能代谢甘露醇的能力,开发了一种稳定的同位素标记方法来跟踪宿主巨噬细胞和上皮细胞细胞内增殖过程中肠道沙门氏菌的代谢产物。通过液相色谱-质谱法(LC-MS)测量沙门氏菌代谢产物中的标签掺入,并将其与代谢建模相结合,我们确定了沙门氏菌使用的相关碳源,揭示了其代谢途径,并量化了中心碳代谢的相对反应率。我们的研究结果强调了Entner-Doudoroff途径(EDP)和磷酸烯醇丙酮酸羧化酶对细胞内增殖的沙门氏菌的重要性。更广泛地说,我们的代谢标记策略为了解病原体在宿主细胞内的代谢开辟了新的途径。
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来源期刊
PLoS Biology
PLoS Biology 生物-生化与分子生物学
CiteScore
14.40
自引率
2.00%
发文量
359
审稿时长
3 months
期刊介绍: PLOS Biology is an open-access, peer-reviewed general biology journal published by PLOS, a nonprofit organization of scientists and physicians dedicated to making the world's scientific and medical literature freely accessible. The journal publishes new articles online weekly, with issues compiled and published monthly. ISSN Numbers: eISSN: 1545-7885 ISSN: 1544-9173
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