Neff John, DeMeester Kristen E, Parraga Paola K, Suciu Radu, Dix Melissa, Simon Gabriel, Gianakopoulos Max A, Melillo Bruno, Cravatt Benjamin F, Shiloh Michael U
Department of Internal Medicine University of Texas Southwestern Medical Center Dallas, TX 75390, USA.
Department of Microbiology, University of Texas Southwestern Medical Center Dallas, TX 75390, USA.
bioRxiv. 2025 Sep 3:2025.08.30.673236. doi: 10.1101/2025.08.30.673236.
Innate immune cells such as monocytes and macrophages provide the earliest defense against infection by intracellular pathogens by initiating signaling pathways and restricting pathogen replication. However, the full complement of proteins that mediate cell-autonomous immunity remains incompletely defined. Here, we applied cysteine-directed activity-based protein profiling (ABPP) to map proteome-wide cysteine reactivity changes in THP-1 monocytes and primary human monocyte-derived macrophages during (Mtb) infection. Across both cell types, we quantified 148 cysteine residues with altered reactivity. Genetic perturbation of a subset of proteins harboring these changes significantly impacted Mtb replication, revealing functional links between site-specific cysteine reactivity and antimicrobial defense. These data define previously unrecognized host protein changes during Mtb infection and provide a resource for investigating post-translational events that regulate innate immune responses to intracellular bacteria.
诸如单核细胞和巨噬细胞等先天免疫细胞通过启动信号通路和限制病原体复制，对细胞内病原体感染提供最早的防御。然而，介导细胞自主免疫的蛋白质的完整组成仍未完全明确。在此，我们应用基于半胱氨酸导向活性的蛋白质谱分析（ABPP）来绘制结核分枝杆菌（Mtb）感染期间THP-1单核细胞和原代人单核细胞衍生巨噬细胞中全蛋白质组半胱氨酸反应性变化图谱。在这两种细胞类型中，我们定量了148个反应性改变的半胱氨酸残基。对具有这些变化的一部分蛋白质进行基因扰动显著影响了Mtb复制，揭示了位点特异性半胱氨酸反应性与抗菌防御之间的功能联系。这些数据定义了Mtb感染期间以前未被认识到的宿主蛋白质变化，并为研究调节对细胞内细菌先天免疫反应的翻译后事件提供了资源。
