细胞内ATP的利用在细胞过程中起着至关重要的作用,这对于调节宿主-病原体动力学和随后的免疫反应至关重要。这项研究的重点是ATP结合蛋白,以剖析金黄色葡萄球菌和人类细胞之间的复杂相互作用,特别是巨噬细胞(THP-1)和角质形成细胞(HaCaT),在细胞内感染期间。使用脱硫生物素-ATP探针提供了各种蛋白质活性和功能的快照,靶向ATP相互作用蛋白。在金黄色葡萄球菌中,我们观察到营养获取所需途径的富集,氨基酸的生物合成和代谢,和能量代谢位于人体细胞内。此外,蛋白质活性的直接分析揭示了金黄色葡萄球菌对角质形成细胞和巨噬细胞的特异性适应。用细胞内细菌将差异激活的蛋白质映射到人细胞中的生化途径,揭示了细胞类型特异性适应细菌挑战,其中THP-1细胞优先考虑免疫防御,自噬性细胞死亡,和炎症。相比之下,HaCaT细胞强调屏障完整性和免疫激活。我们还观察到宿主过程和代谢变化的细菌调节。这些发现为金黄色葡萄球菌-宿主细胞相互作用的动力学提供了有价值的见解,发光调节宿主对金黄色葡萄球菌的免疫反应,这可能涉及开发免疫调节疗法。
目的:这项研究使用化学蛋白质组学方法靶向活性ATP相互作用蛋白,并检查金黄色葡萄球菌与人细胞系THP-1和HaCaT之间的动态蛋白质组学相互作用。它揭示了细菌感染过程中巨噬细胞和角质形成细胞的不同反应。金黄色葡萄球菌表现出对每种细胞类型的细胞内环境的定制响应和在暴露于专业和非专业吞噬细胞期间的适应性。它还强调了金黄色葡萄球菌在宿主细胞内持续使用的策略。这项研究提供了人类细胞对金黄色葡萄球菌感染反应的重要见解,阐明了构成巨噬细胞和角质形成细胞防御机制基础的复杂蛋白质组变化。值得注意的是,这项研究强调了宿主的代谢重编程和免疫策略之间的微妙的相互作用,提出了增强宿主防御和抑制细菌存活的潜在治疗靶点。这些发现增强了我们对宿主-病原体相互作用的理解,并可以为针对金黄色葡萄球菌感染的靶向治疗的发展提供信息。
The utilization of ATP within cells plays a fundamental role in cellular processes that are essential for the regulation of host-pathogen dynamics and the subsequent immune response. This study focuses on ATP-binding proteins to dissect the complex interplay between Staphylococcus aureus and human cells, particularly macrophages (THP-1) and keratinocytes (HaCaT), during an intracellular infection. A snapshot of the various protein activity and function is provided using a desthiobiotin-ATP probe, which targets ATP-interacting proteins. In S. aureus, we observe enrichment in pathways required for nutrient acquisition, biosynthesis and metabolism of amino acids, and energy metabolism when located inside human cells. Additionally, the direct profiling of the protein activity revealed specific adaptations of S. aureus to the keratinocytes and macrophages. Mapping the differentially activated proteins to biochemical pathways in the human cells with intracellular bacteria revealed cell-type-specific adaptations to bacterial challenges where THP-1 cells prioritized immune defenses, autophagic cell death, and inflammation. In contrast, HaCaT cells emphasized barrier integrity and immune activation. We also observe bacterial modulation of host processes and metabolic shifts. These findings offer valuable insights into the dynamics of S. aureus-host cell interactions, shedding light on modulating host immune responses to S. aureus, which could involve developing immunomodulatory therapies.
OBJECTIVE: This study uses a chemoproteomic approach to target active ATP-interacting proteins and examines the dynamic proteomic interactions between Staphylococcus aureus and human cell lines THP-1 and HaCaT. It uncovers the distinct responses of macrophages and keratinocytes during bacterial infection. S. aureus demonstrated a tailored response to the intracellular environment of each cell type and adaptation during exposure to professional and non-professional phagocytes. It also highlights strategies employed by S. aureus to persist within host cells. This study offers significant insights into the human cell response to S. aureus infection, illuminating the complex proteomic shifts that underlie the defense mechanisms of macrophages and keratinocytes. Notably, the study underscores the nuanced interplay between the host\'s metabolic reprogramming and immune strategy, suggesting potential therapeutic targets for enhancing host defense and inhibiting bacterial survival. The findings enhance our understanding of host-pathogen interactions and can inform the development of targeted therapies against S. aureus infections.